{"text_format":"1","workbook":"xlsx","n_sheets":11,"sheets":["Introduction","Conversions","Immobilization","Flowcell","Diffusion","Dilution","Equilibrium","Complex calc.","Solvent calibration","Simulation","Kineticplot"],"not_rendered":{"charts":10,"images":1,"macros":1,"external_links":0,"defined_names":7,"comments":0,"pivot_tables":0},"unopened":[],"bounds":{"document_bytes":16777216,"sheet_cells":100000,"rows_first":2000,"rows_last":500,"rows_between":2000,"halvings":0,"cell_chars":16384,"cell_head_chars":4096},"cells_cut":0,"cells_raw":0,"read":"whole","date_system":"1900"}

{"sheet":"Introduction","index":0,"n_rows":32,"n_cols":14,"rows_kept":32,"cut":false,"hidden":false,"merged":[[22,1,25,2],[3,1,3,2],[3,4,3,6],[12,4,12,6],[18,1,18,2]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Introduction",null,null,null,null,null,null,null,null,null,"Content"]
[]
[null,"The abbreviations used in the formulas are:",null,null,"The units used in the formulas are:"]
[null,"Name","Meaning",null,"Symbol","Name","SI units",null,null,null,null,"Introduction",null,"This page"]
[null,"Da","Dalton",null,"T","Absolute temperature","K (kelvin)",null,null,null,null,"Conversions",null,"Convert ligand immobilization to concentrations and calculate solution concentrations"]
[null,"RU","response units",null,"N","Newton","kg m s-2",null,null,null,null,"Immobilization",null,"Calculate immobilization parameters"]
[null,"M","Molar",null,"Pa","Pascal","N m-2",null,null,null,null,"Flow cell",null,"Calculate flow cell parameters"]
[null,"Mr","molecular mass in Da",null,"J","Joule","N m",null,null,null,null,"Diffusion",null,"Calculate diffusion and mass transport parameters"]
[null,"R","response in RU",null,null,"Liquid viscosity","Poise = Pa s",null,null,null,null,"Dilution",null,"Calculate a dilution scheme - three variants"]
[null,"Req","equilibrium response in RU",null,null,null,null,null,null,null,null,"Equilibrium",null,"Calculate equilibrium and dissociation based on kinetics and analyte concentration"]
[null,"Rmax","maximal response in RU",null,null,null,null,null,null,null,null,"Complex calculation",null,"Calculate complex formation"]
[null,"ka","association rate constant in M-1s-1",null,"The constants used in the formulas are:",null,null,null,null,null,null,"Solvent calibration",null,"Calculate solvent calibration dilution and curve"]
[null,"kd","dissociation rate constant in s-1",null,"Symbol","Name","SI units",null,null,null,null,"Simulation",null,"Simulate association and dissociation curves"]
[null,"KD","equilibrium constant in M",null,"NA","constant of Avogadro","6.0220 1023 mol-1",null,null,null,null,"Kinetic plot",null,"Excel format to make kinetic plot "]
[null,"D","diffusion constant in m2s-1",null,"R","Gas constant","8.3144 J mol-1 K-1"]
[null,null,null,null,"k","constant of Boltzmann","1.3807 10-23 J K-1"]
[null,null,null,null,"h","constant of Planck","6.6262 10-34 J s"]
[null,"Each spreadsheet  is divided in two parts:",null,null,"c","speed of light (vacuum)","2.9979 108 m s-1"]
[null,"yellow","input of data"]
[null,"orange","output of calculations"]
[]
[null,"Please note that in order to obtain useful results, the input values should be in the correct units (as indicated per input cell).",null,null,null,null,null,null,null,null,null,"References"]
[null,null,null,null,null,null,null,null,null,null,null,"Kortt, A. A., Nice, E. and Gruen, L. C.; Analytical Biochemistry (273) 1: 133-141; 1999."]
[null,null,null,null,null,null,null,null,null,null,null,"Walters, R. R., Graham, J. F., Moore, R. M., et al.; Analytical Biochemistry (140) 1: 190-195; 1984."]
[null,null,null,null,null,null,null,null,null,null,null,"Ota, A. and Ueda, S.; Hybridoma (17) 5: 471-477; 1998."]
[]
[null,null,null,null,null,null,null,null,null,null,null,"Biacore AB; BiaSimulation. 1998."]
[null,"The sheets are locked to protect the formulas."]
[null,"Password:","sprpages",null,null,null,null,null,null,null,null,"Go to www.sprpages.nl and SPRpages book for more information."]
[]
[null,"Version 4.0","2022-05-24"]

{"sheet":"Conversions","index":1,"n_rows":26,"n_cols":20,"rows_kept":26,"cut":false,"hidden":false,"merged":[[6,8,6,11],[12,8,12,11]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Conversions",null,null,null,null,null,null,null,null,null,null,null,null,"Conversions"]
[]
[null,"Response / binding sites / concentration",null,null,null,null,null,"Concentration calculation",null,null,null,null,null,null,"Concentration calculation"]
[]
[null,"Input"]
[null,"Response",500,"RU",null,null,null,null,"Stock concentration",null,null,null,null,null,"Compount","Amount","Mw","Amount","Volume","Conc."]
[null,"Molecular mass",60000,"Da",null,null,null,"Protein","(mg mL-1)","(Da)","(M)","(µM)",null,null,null,"(gram)","(Da)","(mol)","(mL)","(mol/L )"]
[null,null,null,null,null,null,null,"A",2.7,150000,{"$f":"I9/J9","v":1.8E-5},{"$f":"K9*1000000","v":18}]
[null,"Calculated",null,null,null,null,null,"B",4.66,150000,{"$f":"I10/J10","v":3.1066666666666665E-5},{"$f":"K10*1000000","v":31.066666666666666},null,null,"T101",8.9599999999999992E-3,25600,{"$f":"P10/Q10","v":3.4999999999999998E-7},1,{"$f":"R10*1000/S10","v":3.5E-4}]
[null,"Binding sites",{"$f":"C7/C8","v":8.3333333333333332E-3},"pmol mm-2"]
[null,"Binding sites",{"$f":"C15/C8","v":8.3333333333333335E-9},"mol m-2"]
[null,null,null,null,null,null,null,null,"Pre dilutions"]
[null,"Surface density",{"$f":"C7/1000","v":0.5},"ng mm-2",null,null,null,null,"Stock","Volume","End vol.","Conc."]
[null,"Surface density",{"$f":"C7/1000000","v":5.0000000000000001E-4},"g m-2",null,null,null,null,{"$f":"L8","v":"(µM)"},"(µl)","(µl)","(nM)"]
[null,null,null,null,null,null,null,{"$f":"H9","v":"A"},{"$f":"L9","v":18},10,{"$f":"(I16*J16)*(1000/L16)","v":720},250]
[null,"Concentration",{"$f":"C7/(100*C8)","v":8.3333333333333331E-5},"mol L-1",null,null,null,{"$f":"H10","v":"B"},{"$f":"L10","v":31.066666666666666},10,{"$f":"(I17*J17)*(1000/L17)","v":310.66666666666669},1000]
[null,"Concentration",{"$f":"C7/100","v":5},"mg mL-1"]
[]
[]
[null,"Response / binding sites / concentrations",null,null,null,null,null,"Concentration calculation and pre-dilution",null,null,null,null,null,null,"Concentration calculation"]
[]
[null,"1) Add response and ligand mass",null,null,null,null,null,"1) Add concentration and molecular weight.",null,null,null,null,null,null,"1) Add concentration and molecular weight."]
[null,"2) Calculated are binding sites / surface density",null,null,null,null,null,"2) Add minimal volume from stock: e.g. 10 µL",null,null,null,null,null,null,"2) Add volume of buffer added"]
[null,"    and volume concentration",null,null,null,null,null,"3) Add end concentration of pre-dilution in nM",null,null,null,null,null,null,"3) See concentration in solution"]
[null,null,null,null,null,null,null,"4) See end volume of pre-dilution"]

{"sheet":"Immobilization","index":2,"n_rows":30,"n_cols":11,"rows_kept":30,"cut":false,"hidden":false,"merged":[]}
["BiaCalculations"]
["Immobilization",null,null,null,null,null,null,null,null,null,"Help"]
[]
[]
[null,"Calculations before immobilization",null,null,null,null,"Calculations after immobilization"]
[]
[null,"Ligand",null,null,null,null,"Ligand"]
[null,"Molecular mass",36500,"Da",null,null,"Molecular mass",36500,"Da"]
[null,"Valency",1,null,null,null,"Immobilization",2000,"RU"]
[null,"Activity",80,"%",null,null,"Valency",1]
[]
[null,"Analyte",null,null,null,null,"Analyte"]
[null,"Molecular mass",2500,"Da",null,null,"Molecular mass",45000,"Da"]
[null,"Desired Rmax",100,"RU",null,null,"Rmax",80,"RU"]
[]
[null,"Calculations",null,null,null,null,"Calculations"]
[null,"Ligand immobilization",{"$f":"((C8*C14)/(C13*C9))/(C10/100)","v":1825},"RU",null,null,"Rmax analyte",{"$f":"H13/H8*H9*H10","v":2465.7534246575342},"RU"]
[null,"Ligand sites",{"$f":"(C17/C8*C9)/(C10/100)","v":6.25E-2},"pmol mm-2",null,null,"Ligand sites",{"$f":"H9/H8*H10","v":5.4794520547945202E-2},"pmol mm-2"]
[null,null,null,null,null,null,"Ligand activity",{"$f":"(H14/H9*(H8/H13)/H10)*100","v":3.2444444444444445},"%"]
[null,null,null,null,null,null,"Ligand concentration",{"$f":"H9/(100*H8)","v":5.4794520547945202E-4},"mol L-1"]
[]
[]
[]
[]
[null,"Calculations before immobilization",null,null,null,null,"Calculations after immobilization"]
[]
[null,"1) Add ligand molecular mass and valency",null,null,null,null,"1) Add ligand data"]
[null,"2) Add activity after immobilization if known ",null,null,null,null,"2) Add analyte molecular mass and observed Rmax"]
[null,"3) Add analyte molecular mas and desired Rmax",null,null,null,null,"3) See calculated values"]
[null,"4) See amount of ligand to be immobilized"]

{"sheet":"Flowcell","index":3,"n_rows":26,"n_cols":21,"rows_kept":26,"cut":false,"hidden":false,"merged":[[25,9,28,14],[3,9,3,14],[3,1,3,6],[13,1,13,6],[13,9,13,14]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Flow and Flow cell",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Flow and Flow cell"]
[]
[null,"Flow rate",null,null,null,null,null,null,null,"Time shift between flow cells",null,null,null,null,null,null,null,"Wall shear"]
[]
[]
[null,"flow rate","f",1,"µL min-1",null,null,null,null,"flow","f",10,"µL min-1",null,null,null,null,"flowrate","q",50,"µL min-1"]
[null,null,null,null,null,null,null,null,null,"volume","v",0.3,"µL min-1",null,null,null,null,"wall shear","jw",{"$f":"T7/T9","v":4132.2314049586776},"s-1"]
[null,null,null,{"$f":"(D$7/60)*0.000001","v":1.6666666666666667E-8},"L s-1",null,null,null,null,null,null,null,null,null,null,null,null,"factor","f",1.21E-2]
[null,null,null,{"$f":"(D$7/60)*0.000000001","v":1.6666666666666667E-11},"m3 s-1",null,null,null,null,"shift",null,{"$f":"(L8/L7)*60","v":1.7999999999999998},"s"]
[]
[]
[]
[null,"Flow cell (Biacore 2000/3000)",null,null,null,null,null,null,null,"Thickness of diffusion layer"]
[]
[]
[null,"flow rate","f",3,"µL min-1",{"$f":"((D17/60)*0.000000001)","v":5.0000000000000008E-11},"m3 s-1",null,null,"Diffusion factor","D",5.0000000000000002E-11,"m2 s-1",{"$f":"L17","v":5.0000000000000002E-11},"m2 s-1"]
[null,"height","h",0.05,"mm",{"$f":"D18/1000","v":5.0000000000000002E-5},"m",null,null,"flow rate","f",3,"µL min-1",{"$f":"((L18/60)*0.000000001)","v":5.0000000000000008E-11},"m3 s-1"]
[null,"width","w",0.3,"mm",{"$f":"D19/1000","v":2.9999999999999997E-4},"m",null,null,"height","h",0.05,"mm",{"$f":"L19/1000","v":5.0000000000000002E-5},"m"]
[null,"length","l",0.8,"mm",{"$f":"D20/1000","v":8.0000000000000004E-4},"m",null,null,"width","w",0.3,"mm",{"$f":"L20/1000","v":2.9999999999999997E-4},"m"]
[null,null,null,null,null,null,null,null,null,"length","l",0.8,"mm",{"$f":"L21/1000","v":8.0000000000000004E-4},"m"]
[null,"volume","v",{"$f":"D18*D19*D20","v":1.2E-2},"mm3",{"$f":"F18*F19*F20","v":1.1999999999999999E-11},"m3"]
[null,"refresh rate",null,{"$f":"D17/D22","v":250},"times min-1",{"$f":"F17/F22","v":4.1666666666666679},"times s-1",null,null,"thickness","d",{"$f":"N23*1000000","v":8.4343266530175107},"µm",{"$f":"((N17*N19^2*N20*N21)/(N18))^(1/3)","v":8.4343266530175106E-6},"m"]
[]
[]
[null,null,null,null,null,null,null,null,null,"Glaser, R. W.; Antigen-antibody binding and mass transport by convection and diffusion to a surface: a two-dimensional computer model of binding and dissociation kinetics. Analytical Biochemistry (213) 1: 152-161; 1993."]

{"sheet":"Diffusion","index":4,"n_rows":32,"n_cols":19,"rows_kept":32,"cut":false,"hidden":false,"merged":[[3,15,3,19],[17,15,17,19],[3,1,3,5],[3,7,3,11],[5,7,5,10],[17,1,17,5],[17,7,17,11]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Diffusion",null,null,null,null,null,null,null,null,null,null,null,null,null,"Diffusion"]
[]
[null,"Diffusion factor for globular protein",null,null,null,null,null,"Dynamic viscosity of water",null,null,null,null,null,null,null,"Diffusion coefficients of some proteins"]
[]
[null,null,null,null,null,null,null,"v = A x 10^(B/(T-C)"]
[null,"molecular mass","Mr",178000,"Da",null,null,null,null,null,null,null,null,null,null,"Protein","Mw","D20,w"]
[null,"friction factor","f/fo",1.3,null,null,null,null,"A",2.4139999999999999E-5,"Pa.s",null,null,null,null,null,"(Da)","(x 10-11 m2s-1)"]
[null,"solution viscosity","v/vo",0.89,null,null,null,null,"B",247.8,"K",null,null,null,null,"Albumin, bovine serum",66500,"6.29 ± 0.10"]
[null,"factor 1",null,9.9999999999999994E-12,null,null,null,null,"C",140,"K",null,null,null,null,"Albumin, human serum",69000,"6.43 ± 0.07"]
[null,"factor 2",null,324.3,null,null,null,"Temperature","T",{"$f":"273+20","v":293},"K",null,null,null,null,"IgG, human",153000,"4.11 ± 0.05"]
[]
[null,"Diffusion factor","D",{"$f":"((D10*D11)*(D7^(-(0.333))))/(D8*D9)","v":5.0029219660526572E-11},"m2 s-1",null,null,"Dynamic viscosity","v",{"$f":"J8*10^(J9/(J11-J10))","v":1.0054144180571549E-3},"Pa.s",null,null,null,null,"Ref: (211)"]
[]
[]
[]
[]
[null,"Mass transport coefficient (km)",null,null,null,null,null,"Mass transport coefficient (kt)",null,null,null,null,null,null,null,"Dynamic viscosity of water"]
[]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"The dynamic viscosity of water is 8.90 × 10−4 Pa·s or 8.90 × 10−3 dyn·s/cm2 or 0.890 cP at about 25 °C."]
[null,"Diffusion factor","D",{"$f":"D13","v":5.0029219660526572E-11},"m2 s-1",null,null,"km",null,{"$f":"D29","v":8.6817590426761711E-6},"m s-1",null,null,null,null,"Temperature ","Viscosity ",null,"Water has a viscosity of 0.0091 poise at 25 °C, or 1 centipoise at 20 °C."]
[null,"flow rate","f",{"$f":"Flowcell!F17","v":5.0000000000000008E-11},"m/s",null,null,"Molecular weight","Mr",{"$f":"D7","v":178000},"Da",null,null,null,null,"[°C]","[mPa·s]",null,"As a function of temperature T (K): (Pa·s) = A × 10B/(T−C)"]
[null,"flow cell height","h",{"$f":"Flowcell!F18","v":5.0000000000000002E-5},"m",null,null,"Factor",null,1000000000,null,null,null,null,null,10,1.3080000000000001,null,"where A=2.414 × 10−5 Pa·s ; B = 247.8 K ; and C = 140 K.[citation needed]"]
[null,"flow cell width","w",{"$f":"Flowcell!F19","v":2.9999999999999997E-4},"m",null,null,null,null,null,null,null,null,null,null,20,1.002]
[null,"flow cell length","l",{"$f":"Flowcell!F20","v":8.0000000000000004E-4},"m",null,null,null,null,null,null,null,null,null,null,30,0.79779999999999995]
[null,"factor 1",null,0.3,null,null,null,null,null,null,null,null,null,null,null,40,0.65310000000000001]
[null,"factor 2",null,0.98,null,null,null,null,null,null,null,null,null,null,null,50,0.54710000000000003]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,60,0.46579999999999999]
[null,"km",null,{"$f":"D27*((((D21)^2*D22)/(D26*(D23^2)*D24*D25))^(1/3))","v":8.6817590426761711E-6},"m s-1",null,null,"kt",null,{"$f":"J21*J22*J23","v":1545353109.5963583},"RU M-1 s-1",null,null,null,null,70,0.40439999999999998]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,80,0.35499999999999998]
[null,"Values come from flowcell sheet",null,null,null,null,null,"The kt value is 10 times higher than in publication ref32",null,null,null,null,null,null,null,90,0.315]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,100,0.28220000000000001]

{"sheet":"Dilution","index":5,"n_rows":40,"n_cols":48,"rows_kept":40,"cut":false,"hidden":false,"merged":[[21,47,23,51],[4,47,15,51],[9,37,9,38],[10,37,10,38],[6,44,6,45],[4,37,4,40],[4,42,4,45],[7,37,7,38],[8,37,8,38],[6,42,6,43],[7,42,7,43],[8,42,8,43],[9,42,9,43],[14,36,14,38],[19,40,19,41],[19,42,19,44],[13,42,13,43],[19,37,19,39],[5,5,5,6],[6,5,6,6],[4,29,15,34],[18,1,18,2],[18,5,18,6],[4,5,4,6],[10,42,10,43],[13,36,13,38],[15,36,15,38],[6,39,6,40],[4,11,15,16],[13,3,13,4],[4,25,4,26],[4,7,4,8],[7,5,7,6],[8,5,8,6],[18,19,18,20],[18,23,18,24],[4,23,4,24],[5,23,5,24],[6,23,6,24],[7,23,7,24],[8,23,8,24],[13,21,13,22]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Dilution wizard",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Dilution wizard with duplo sample",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Dilution wizard for competition experiments"]
[]
[null,null,"Biacore 2000/3000","T100/200",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Biacore 2000/3000","T100/200"]
[null,"Inject command","KInject","Inject",null,"Protein",null,null,null,null,null,"1) Select Injection type and flow rate\n2) Select injection time and replicates\n\n3) Select dilution step and adjust final volume\n4) Select highest injection concentration and stock concentration\n\nOn the row of vial 1 is the first dilution from the stock\nNext vials are diluted from the previous vial\n\n5) Add kinetic parameters to calculate theorectical Req\n6) Select the stock glycerol concentration to calculate the final concentration in the samples",null,null,null,null,null,null,null,"Inject command","KInject","Inject",null,"Protein",null,null,null,null,null,"1) Select Injection type and flow rate\n2) Select injection time and replicates\n\n3) Select dilution step and adjust final volume\n4) Select highest injection concentration and stock concentration\n\nOn the row of vial 1 is the first dilution from the stock\nNext vials are diluted from the previous vial\n\n5) Add kinetic parameters to calculate theorectical Req\n6) Select the stock glycerol concentration to calculate the final concentration in the samples",null,null,null,null,null,null,null,"Constant concentration",null,null,null,null,"Competition concentration",null,null,null,null,"1) Select number of dilutions, final volume, constant concentration and stock concentration\n2) Select dilution step, highest concentration and stock concentration\n\n3) Prepare first constant dilution and add volume in vial 1\n4) Dilute const. conc. in vials 0 (no competitor) and vials 2 → n (see table)\n\n5) Dilute comp. concentration (first dilution) and add to first vial\n6) Dilute the same volume from vial 1 to vial 2 etc."]
[null,"Flow rate",30,30,"ul/min","Dilution step",null,2,"x",null,null,null,null,null,null,null,null,null,null,"Flow rate",30,30,"ul/min","Dilution step",null,2,"x"]
[null,"Inject time",5,2,"min","Final volume",null,125,"µL",null,null,null,null,null,null,null,null,null,null,"Inject time",5,3,"min","Final volume",null,210,"µL",null,null,null,null,null,null,null,null,null,null,null,"Protein",null,null,null,"Protein"]
[null,"Replicates",1,2,"x","Highest conc.",null,500,"nM",null,null,null,null,null,null,null,null,null,null,"Replicates",1,1,"x","Highest conc.",null,100,"uM",null,null,null,null,null,null,null,null,null,null,"no. of dilutions + 1",null,7,null,null,"Dilution step",null,2,"x"]
[null,"Extra volume",{"$f":"LOOKUP(C5,B12:B14,C12:C14)","v":40},40,"ul","Stock concentration",null,4333,{"$f":"I8","v":"nM"},null,null,null,null,null,null,null,null,null,null,"Extra volume",{"$f":"LOOKUP(U5,T12:T14,U12:U14)","v":40},58,"ul","Stock concentration",null,4333,{"$f":"AA8","v":"uM"},null,null,null,null,null,null,null,null,null,null,"Final volume",null,125,"µL",null,null,null,{"$f":"AN9","v":125}]
[null,"Total volume",{"$f":"((C6*C7)+C9)*C8","v":190},{"$f":"((D6*D7)+D9)*D8","v":200},"ul",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Total volume",{"$f":"((U6*U7)+U9)*U8","v":190},{"$f":"((V6*V7)+V9)*V8","v":148},"ul",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Constant conc.",null,125,"nM",null,"Highest conc.",null,500,"uM"]
[null,null,"    ",null,null,null,"ka",500000,"M-1s-1",null,null,null,null,null,null,null,null,null,null,null,"    ",null,null,null,"ka",10000,"M-1s-1",null,null,null,null,null,null,null,null,null,null,"Stock concentration",null,61600,{"$f":"AO10","v":"nM"},null,"Stock concentration",null,4333,{"$f":"AT10","v":"uM"}]
[null,"Inject",30,"max volume injection",null,null,"kd",5.0000000000000001E-4,"s-1",null,null,null,null,null,null,null,null,null,null,"Inject",30,"max volume injection",null,null,"kd",1E-3,"s-1",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"extra"]
[null,"KInject",40,250,"µL",null,"Solvent",0.01,"%",null,null,null,null,null,null,null,null,null,null,"KInject",40,250,"µL",null,"Solvent",600,"%",null,null,null,null,null,null,null,null,null,null,null,null,"stock ","buffer",1.1000000000000001,null,null,"stock ","buffer"]
[null,"Quickinject",10,"Check volume",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Quickinject",10,"Check volume",null,null,null,null,null,null,null,null,null,null,null,null,null,null,{"$f":"CONCATENATE(\"First dilution (\",AM24,\" nM)\")","v":"First dilution (250 nM)"},null,null,{"$f":"AP13*((AN8-1)*AN9*AN10+(AN9*2*AN10))/AN11","v":2.2321428571428572},{"$f":"(AN9+((AN8-1)*AN9/2))*AP13","v":550},{"$f":"CONCATENATE(AN9,\" in vial 1\")","v":"125 in vial 1"},{"$f":"CONCATENATE(\"First dilution (\",AP24,\" \",AT10,\")\")","v":"First dilution (1000 uM)"},null,{"$f":"AN9*AS10*2*AP13/AS11","v":31.733210246942072},{"$f":"AN9*AP13-AS14","v":105.76678975305794}]
[null,null,null,null,null,null,"KD",{"$f":"H12/H11","v":1.0000000000000001E-9},"M",null,null,null,null,null,null,null,null,null,null,"Factor",1,null,null,null,"KD",{"$f":"Z12/Z11","v":9.9999999999999995E-8},"M",null,null,null,null,null,null,null,null,null,"No competitor",null,null,{"$f":"AN16","v":62.5},{"$f":"AO16","v":62.5},"vial 0"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Second → n dilution",null,null,{"$f":"AN9/2","v":62.5},{"$f":"AN16","v":62.5},"vial 2 → n"]
[null,"Dilution scheme",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Dilution scheme"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Dilution scheme"]
[null,"Stock",null,"buffer","mix","Final solution",null,"KD","Req","Solvent",null,null,null,null,null,null,null,null,null,"Stock",null,"buffer","mix","Final solution",null,"KD","Req","Solvent"]
["Vial",{"$f":"CONCATENATE(\"(\",I8,\")\")","v":"(nM)"},"(µL)","(µL)","(µL)",{"$f":"CONCATENATE(\"(\",I8,\")\")","v":"(nM)"},"(µL)","(x)","(%)",{"$f":"CONCATENATE(\"(\",I13,\")\")","v":"(%)"},null,null,null,null,null,null,null,null,"Vial",{"$f":"CONCATENATE(\"(\",AA8,\")\")","v":"(uM)"},"(µL)","(µL)","(µL)",{"$f":"CONCATENATE(\"(\",AA8,\")\")","v":"(uM)"},"(µL)","(x)","(%)",{"$f":"CONCATENATE(\"(\",AA13,\")\")","v":"(%)"},null,null,null,null,null,null,null,null,null,"Constant conc.",null,null,"Competitor conc.",null,"Final volume and concentrations"]
[1,{"$f":"H9","v":4333},{"$f":"(H6/(H6-1)*H7)/B21*H8","v":28.848372951765523},{"$f":"(H6/(H6-1)*H7)-C21","v":221.15162704823447},{"$f":"SUM(C21:D21)","v":250},{"$f":"C21*B21/E21","v":500.00000000000006},{"$f":"E21-C22","v":125},{"$f":"(F21/1000000000)/(H$12/H$11)","v":500.00000000000006},{"$f":"(((H$11*(F21/C$39))/(H$11*(F21/C$39)+H$12)))*100","v":99.800399201596804},{"$f":"C21*H13/E21","v":1.153934918070621E-3},null,null,null,null,null,null,null,null,1,{"$f":"Z9","v":4333},{"$f":"W21*T22/T21","v":12.116316639741518},{"$f":"W21-U21","v":512.88368336025849},{"$f":"(Z7+U22)*U15","v":525},{"$f":"U21*T21/W21","v":100},{"$f":"W21-U22","v":210},{"$f":"(X21/1000000000)/(Z$12/Z$11)","v":1},{"$f":"(((Z$11*(X21/U$39))/(Z$11*(X21/U$39)+Z$12)))*100","v":99.900099900099917},{"$f":"U21*Z13/W21","v":13.84721901684745},null,null,null,null,null,null,null,null,null,"protein","conc.","buffer","protein","conc.","volume","const.","variabl."]
[2,{"$f":"F21","v":500.00000000000006},{"$f":"(1/H$6)*E21","v":125},{"$f":"(C22*H$6)-C22","v":125},250,{"$f":"C22*F21/E22","v":250.00000000000003},125,250.00000000000003,99.601593625498012,{"$f":"J21/H$6","v":5.7696745903531048E-4},null,null,null,null,null,null,null,null,2,{"$f":"Z8","v":100},{"$f":"W22*T23/T22","v":315},{"$f":"W22-U22","v":315},{"$f":"(2*Z7+U23)*U15","v":630},{"$f":"U22*X21/W22","v":50},{"$f":"W22-U23","v":420},0.5,99.800399201596804,{"$f":"AB21/Z$6","v":6.9236095084237252},null,"ßduplo sample",null,null,null,null,null,null,"Vial","(µL)",{"$f":"CONCATENATE(\"(\",AO10,\")\")","v":"(nM)"},"(µL)","(µL)",{"$f":"CONCATENATE(\"(\",AT10,\")\")","v":"(uM)"},"(µL)",{"$f":"CONCATENATE(\"(\",AO10,\")\")","v":"(nM)"},{"$f":"CONCATENATE(\"(\",AT10,\")\")","v":"(uM)"},null,null,"Don't forget to make a sample without competitor"]
[3,250.00000000000003,125,125,250,125.00000000000001,125,125.00000000000001,99.206349206349216,2.8848372951765524E-4,null,null,null,null,null,null,null,null,3,{"$f":"T22/Z$6","v":50},{"$f":"Z$7/(Z$6-1)","v":210},{"$f":"Z$7","v":210},{"$f":"SUM(U23:V23)","v":420},25,210,0.25,99.601593625498012,3.4618047542118626,null,null,null,null,null,null,null,null,0,{"$f":"AL25","v":62.5},{"$f":"AM25","v":250},{"$f":"AN25","v":62.5},0,0,{"$f":"AL24+AO24-AO25","v":125},{"$f":"AR24","v":125},0]
[4,125.00000000000001,125,125,250,62.500000000000007,125,62.500000000000007,98.425196850393704,1.4424186475882762E-4,null,null,null,null,null,null,null,null,4,{"$f":"T23/Z$6","v":25},{"$f":"Z$7/(Z$6-1)","v":210},{"$f":"Z$7","v":210},{"$f":"SUM(U24:V24)","v":420},12.5,210,0.125,99.206349206349216,1.7309023771059313,null,null,null,null,null,null,null,null,1,{"$f":"AN9","v":125},{"$f":"AN10*2","v":250},"-",{"$f":"AS9","v":125},{"$f":"AS10*2","v":1000},{"$f":"AL24+AO24-AO25","v":125},{"$f":"AL24*AM24/(AL24+AO24)","v":125},{"$f":"(AO24*AP24)/(AL24+AO24)","v":500}]
[5,62.500000000000007,125,125,250,31.250000000000004,125,31.250000000000004,96.899224806201545,7.2120932379413809E-5,null,null,null,null,null,null,null,null,5,12.5,210,210,420,6.25,210,6.25E-2,98.425196850393704,0.86545118855296566,null,null,null,null,null,null,null,null,2,{"$f":"AL24/2","v":62.5},{"$f":"AM24","v":250},{"$f":"AL25","v":62.5},{"$f":"AO24*(1/(AS8-1))","v":125},{"$f":"AS24","v":500},{"$f":"AL25+AN25+AO25-AO26","v":125},{"$f":"(AL25*AM25+AO24*AR24)/(AL25+AN25+AO24)","v":125},{"$f":"(AO25*AP25)/(AL25+AN25+AO25)","v":250}]
[6,31.250000000000004,125,125,250,15.625000000000002,125,15.625000000000002,93.984962406015043,3.6060466189706905E-5,null,null,null,null,null,null,null,null,6,6.25,210,210,420,3.125,210,3.125E-2,96.899224806201545,0.43272559427648283,null,null,null,null,null,null,null,null,3,{"$f":"AL25","v":62.5},{"$f":"AM25","v":250},62.5,{"$f":"AO25","v":125},{"$f":"AS25","v":250},125,125,{"$f":"(AO26*AP26)/(AL26+AN26+AO26)","v":125}]
[7,15.625000000000002,125,125,250,7.8125000000000009,125,7.8125000000000009,88.652482269503551,1.8030233094853452E-5,null,null,null,null,null,null,null,null,7,3.125,210,210,420,1.5625,210,1.5625E-2,93.984962406015043,0.21636279713824141,null,null,null,null,null,null,null,null,4,62.5,250,62.5,125,125,125,125,62.5]
[8,7.8125000000000009,125,125,250,3.9062500000000004,125,3.9062500000000004,79.617834394904463,9.0151165474267262E-6,null,null,null,null,null,null,null,null,8,1.5625,210,210,420,0.78125,210,7.8125E-3,88.652482269503537,0.10818139856912071,null,null,null,null,null,null,null,null,5,62.5,250,62.5,125,62.5,125,125,{"$f":"(AO28*AP28)/(AL28+AN28+AO28)","v":31.25}]
[9,3.9062500000000004,125,125,250,1.9531250000000002,125,1.9531250000000002,66.137566137566139,4.5075582737133631E-6,null,null,null,null,null,null,null,null,9,0.78125,210,210,420,0.390625,210,3.90625E-3,79.617834394904463,5.4090699284560353E-2,null,null,null,null,null,null,null,null,6,62.5,250,62.5,125,31.25,125,125,15.625]
[10,1.9531250000000002,125,125,250,0.97656250000000011,250,0.97656250000000011,49.407114624505937,2.2537791368566815E-6,null,null,null,null,null,null,null,null,10,0.390625,210,210,420,0.1953125,420,1.953125E-3,66.137566137566139,2.7045349642280177E-2,null,null,null,null,null,null,null,null,7,62.5,250,62.5,125,15.625,125,125,7.8125]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,8,62.5,250,62.5,125,7.8125,125,125,3.90625]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,9,{"$f":"AL31","v":62.5},250,62.5,125,3.90625,125,125,1.953125]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,10,62.5,250,62.5,125,{"$f":"AS32","v":1.953125},{"$f":"AL33+AN33+AO33","v":250},125,0.9765625]
[]
[]
[null,"mM",1000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"mM",1000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"mM"]
[null,"nM",1000000000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"nM",1000000000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"nM"]
[null,"uM",1000000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"uM",1000000,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"uM"]
[null,null,{"$f":"LOOKUP(I8,B36:B38,C36:C38)","v":1000000000},null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,{"$f":"LOOKUP(AA8,T36:T38,U36:U38)","v":1000000},null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"ug/mL"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"mg/mL"]

{"sheet":"Equilibrium","index":6,"n_rows":66,"n_cols":32,"rows_kept":66,"cut":false,"hidden":false,"merged":[]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Equilibrium and dissociation",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Equilibrium and dissociation"]
[]
[null,"Kinetics",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Complex half life",null,null,null,null,null,"Time to 5% dissociation",null,null,null,null,"Time to 95% dissociation"]
[]
[]
[null,"ka",500000,"M-1s-1",null,"KD",{"$f":"C8/C7","v":1.0000000000000001E-9},"M",null,null,null,null,null,null,null,null,null,null,"kd","t1/2","t1/2","t1/2",null,null,"kd","Time","Time",null,null,"kd","Time","Time"]
[null,"kd",5.0000000000000001E-4,"s-1",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"(s-1)","(s)","(min)","(hour)",null,null,"(s-1)","(min)","(hour)",null,null,"(s-1)","(min)","(hour)"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,0.1,{"$f":"LN(2)/S9","v":6.9314718055994522},{"$f":"T9/60","v":0.11552453009332421},{"$f":"U9/60","v":1.9254088348887369E-3},null,null,0.1,{"$f":"(LN(100/95)/Y9)/60","v":8.5488823979250807E-3},{"$f":"Z9/60","v":1.4248137329875133E-4},null,null,0.1,{"$f":"(LN(100/5)/AD9)/60","v":0.49928871225899846},{"$f":"AE9/60","v":8.3214785376499751E-3}]
[null,"Rmax",100,"RU",null,null,null,null,null,null,null,null,null,null,null,null,null,null,0.01,69.314718055994533,1.1552453009332422,1.9254088348887369E-2,null,null,0.01,8.5488823979250797E-2,1.4248137329875134E-3,null,null,0.01,4.9928871225899849,8.3214785376499747E-2]
[null,"Rmax",95,"%",null,null,null,null,null,null,null,null,null,null,null,null,null,null,1E-3,693.14718055994524,11.552453009332421,0.19254088348887369,null,null,1E-3,0.85488823979250805,1.4248137329875133E-2,null,null,1E-3,49.92887122589984,0.83214785376499734]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,1E-4,6931.4718055994526,115.52453009332422,1.925408834888737,null,null,1E-4,8.5488823979250785,0.1424813732987513,null,null,1E-4,499.28871225899843,8.3214785376499734]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,1.0000000000000001E-5,69314.718055994526,1155.2453009332421,19.254088348887368,null,null,1.0000000000000001E-5,85.488823979250796,1.4248137329875132,null,null,1.0000000000000001E-5,4992.8871225899838,83.214785376499734]
[null,"Theoretical Req",null,null,null,null,null,"Time to equilibrium",null,null,null,null,"Complex half life",null,null,null,null,null,9.9999999999999995E-7,693147.18055994529,11552.453009332421,192.54088348887367,null,null,9.9999999999999995E-7,854.8882397925081,14.248137329875135,null,null,9.9999999999999995E-7,49928.871225899849,832.14785376499754]
[]
[null,"C","C","KD","Rmax","Rmax",null,"C","Time","Time","Time",null,"kd","t1/2","t1/2","t1/2"]
[null,"(M)","(nM)","(x)","(RU)","(%)",null,"(M)","(s)","(min)","(hour)",null,"(s-1)","(s)","(min)","(hour)"]
[null,{"$f":"Dilution!F21/1000000000","v":5.0000000000000008E-7},{"$f":"B18*1000000000","v":500.00000000000006},{"$f":"B18/G$7","v":500.00000000000006},{"$f":"(($C$7*B18)/($C$7*B18+$C$8))*$C$10","v":99.800399201596804},{"$f":"(E18/C$10)*100","v":99.800399201596804},null,{"$f":"B18","v":5.0000000000000008E-7},{"$f":"(-LN(1-(C$11/100)))/((C$7*H18)+C$8)","v":11.959011072071814},{"$f":"I18/60","v":0.19931685120119688},{"$f":"J18/60","v":3.3219475200199481E-3},null,{"$f":"C8","v":5.0000000000000001E-4},{"$f":"LN(2)/M18","v":1386.2943611198905},{"$f":"N18/60","v":23.104906018664842},{"$f":"O18/60","v":0.38508176697774738}]
[null,{"$f":"Dilution!F22/1000000000","v":2.5000000000000004E-7},{"$f":"B19*1000000000","v":250.00000000000003},{"$f":"B19/G$7","v":250.00000000000003},{"$f":"(($C$7*B19)/($C$7*B19+$C$8))*$C$10","v":99.601593625498012},99.601593625498012,null,{"$f":"B19","v":2.5000000000000004E-7},23.870376681705093,{"$f":"I19/60","v":0.39783961136175156},6.6306601893625257E-3]
[null,{"$f":"Dilution!F23/1000000000","v":1.2500000000000002E-7},125.00000000000001,125.00000000000001,99.206349206349216,{"$f":"(E20/C$10)*100","v":99.206349206349216},null,1.2500000000000002E-7,47.551305929428402,0.79252176549047337,1.3208696091507889E-2]
[null,{"$f":"Dilution!F24/1000000000","v":6.250000000000001E-8},62.500000000000007,62.500000000000007,98.425196850393704,98.425196850393704,null,6.250000000000001E-8,{"$f":"(-LN(1-(C$11/100)))/((C$7*H21)+C$8)","v":94.353772395401236},{"$f":"I21/60","v":1.5725628732566872},2.6209381220944787E-2,null,"Time to 5% dissociation"]
[null,{"$f":"Dilution!F25/1000000000","v":3.1250000000000005E-8},31.250000000000004,31.250000000000004,96.899224806201545,96.899224806201545,null,3.1250000000000005E-8,185.78184642195282,3.0963641070325472,5.1606068450542456E-2,null,"kd","Time","Time","R"]
[null,{"$f":"Dilution!F26/1000000000","v":1.5625000000000003E-8},15.625000000000002,15.625000000000002,93.984962406015043,93.984962406015043,null,1.5625000000000003E-8,360.38884493882574,6.0064807489804286,0.10010801248300714,null,"(s-1)","(min)","(hour)","(RU)"]
[null,{"$f":"Dilution!F27/1000000000","v":7.8125000000000013E-9},7.8125000000000009,7.8125000000000009,88.652482269503551,88.652482269503551,null,7.8125000000000013E-9,679.88250179948704,11.331375029991451,0.18885625049985752,null,{"$f":"C8","v":5.0000000000000001E-4},{"$f":"(LN(100/95)/M24)/60","v":1.7097764795850161},{"$f":"N24/60","v":2.8496274659750267E-2},{"$f":"C10*(EXP(-(M24*N24*60)))","v":95}]
[null,{"$f":"Dilution!F28/1000000000","v":3.9062500000000007E-9},3.9062500000000004,3.9062500000000004,79.617834394904463,79.617834394904463,null,{"$f":"B25","v":3.9062500000000007E-9},1221.1902261621358,20.353170436035597,0.33921950726725997]
[null,{"$f":"Dilution!F29/1000000000","v":1.9531250000000003E-9},1.9531250000000002,1.9531250000000002,66.137566137566139,66.137566137566139,null,{"$f":"B26","v":1.9531250000000003E-9},2028.8557196556119,{"$f":"I26/60","v":33.8142619942602},{"$f":"J26/60","v":0.56357103323766999}]
[null,{"$f":"Dilution!F30/1000000000","v":9.7656250000000016E-10},0.97656250000000011,0.97656250000000011,49.407114624505937,49.407114624505937,null,{"$f":"B27","v":9.7656250000000016E-10},3031.2547906317054,{"$f":"I27/60","v":50.520913177195091},{"$f":"J27/60","v":0.84201521961991821},null,"Time to 95% dissociation"]
[null,"Use concentrations between 10 - 90% Rmax",null,null,null,null,null,null,null,null,null,null,"kd","Time","Time","R"]
[null,null,null,null,null,null,null,null,null,null,null,null,"(s-1)","(min)","(hour)","(RU)"]
[null,"The concentrations are copied from the dilution page.",null,null,null,null,null,null,null,null,null,null,{"$f":"C8","v":5.0000000000000001E-4},{"$f":"(LN(100/5)/M30)/60","v":99.85774245179968},{"$f":"N30/60","v":1.6642957075299947},{"$f":"C10*(EXP(-(M30*N30*60)))","v":5.0000000000000027}]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[null,"Kinetics"]
[]
[]
[null,"ka",{"$f":"C7","v":500000},"M-1s-1",null,"KD",{"$f":"G7","v":1.0000000000000001E-9},"M"]
[null,"kd",{"$f":"C8","v":5.0000000000000001E-4},"s-1"]
[]
[null,"Rmax",{"$f":"C10","v":100},"RU"]
[null,"Rmax",{"$f":"C11","v":95},"%"]
[]
[null,"Kinetics copied from above."]

{"sheet":"Complex calc.","index":7,"n_rows":100,"n_cols":25,"rows_kept":100,"cut":false,"hidden":false,"merged":[[5,5,8,9],[10,3,10,5],[44,8,51,13],[51,3,51,5]]}
["BiaCalculations"]
["Complex calculations"]
[]
[null,"When [Lo], [Ao] and KD are known",null,null,null,null,null,null,null,"[LA] = (([Lo] + [Ao] + KD) - (([Lo] + [Ao] + KD)^2 - 4*[Lo]*[Ao])^0.5)/2",null,null,null,null,null,null,"At equilibrium the amount of complex is constant",null,null,null,null,null,null,null,"[LA] = [L] * [A] * KD"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,1,"L0 = L + LA"]
[null,"Ligand",2," µM",null,"1) Add concentration constant component\n2) Add highest concentration variable component\n3) Add equilibrium dissocation constant\n4) Add dilution step for variable component",null,null,null,null,null,"[LA] = complex",null,null,null,null,2,"A0 = A + LA"]
[null,"Analyte",10," µM",null,null,null,null,null,null,null,"[Lo] = ligand = constant"]
[null,"KD",3.4999999999999999E-9," (M-1)",null,null,null,null,null,null,null,"[Ao] = analyte = variabel",null,null,null,null,"There are two situations when calculation the complex between two molecules."]
[null,"Dilution",2," (x)",null,null,null,null,null,null,null,null,null,null,null,null,1,"[Ligand] =< KD"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,2,"[Ligand] >> KD"]
[null,"Ligand","Analyte","Complex"]
[null,"FXI","FIX",{"$f":"CONCATENATE(B12,\"_\",C12)","v":"FXI_FIX"},"Sat.",{"$f":"CONCATENATE(B12,\":\",C12)","v":"FXI:FIX"},null,null,null,null,null,null,null,null,null,null,"A","L0 >> A0","then L = L0",null,"ML = M0.L / (KD + L)",null,null,"if L0 >> M0 then ML = M0.L0/(KD + L)"]
["n","(µM)","(µM)","(µM)","(%)","(1:x)"]
[1,{"$f":"C6","v":2},{"$f":"C7","v":10},{"$f":"G27*1000000","v":1.9991254782018339},{"$f":"D14/B14*100","v":99.956273910091696},{"$f":"C14/B14","v":5},null,null,null,null,null,null,null,null,null,null,null,"when dividing by M0 the fractional saturaton of A is calculated by",null,null,null,null,null,null,"Y = [ML]/[M0] = L/(KD + L)"]
[2,{"$f":"B14","v":2},{"$f":"C14/C$9","v":5},1.997671191393217,{"$f":"D15/B15*100","v":99.883559569660846},{"$f":"C15/B15","v":2.5}]
[3,{"$f":"B15","v":2},2.5,1.9864613600245953,99.323068001229757,1.25,null,null,null,null,null,null,null,null,null,null,"B","L0 = ?",null,null,"KD = ([M][L])/[ML] = [M0-ML][L0-ML]/[ML]"]
[4,2,1.25,1.2442378269910874,62.211891349554371,0.625,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"ML = {(L0+M0+KD) - ((L0+M0+KD)^2 - 4 M0L0)^1/2}/2"]
[5,2,0.625,0.62341495275644865,31.170747637822434,0.3125]
[6,2,0.3125,0.31185344059081599,15.5926720295408,0.15625]
[7,2,0.15625,0.15595399925101827,7.7976999625509134,7.8125E-2]
[8,2,7.8125E-2,7.7982992685101699E-2,3.8991496342550849,3.90625E-2]
[9,2,3.90625E-2,3.8992905570747062E-2,1.949645278537353,1.953125E-2]
[10,2,1.953125E-2,1.9496794725523528E-2,0.97483973627617637,9.765625E-3]
[]
[null,"Calculations in steps in M"]
[null,"[L]","[A]","L+A+KD","(L+A+KD)^2","4*L*A","[LA]"]
[1,{"$f":"B14/1000000","v":1.9999999999999999E-6},1.0000000000000001E-5,{"$f":"B27+C27+C$8","v":1.20035E-5},{"$f":"(D27)^2","v":1.4408401224999998E-10},{"$f":"4*B27*C27","v":8.0000000000000008E-11},{"$f":"(D27-(E27-F27)^0.5)/2","v":1.9991254782018339E-6}]
[2,1.9999999999999999E-6,5.0000000000000004E-6,7.0035000000000008E-6,{"$f":"(D28)^2","v":4.9049012250000011E-11},{"$f":"4*B28*C28","v":4.0000000000000004E-11},1.997671191393217E-6]
[3,1.9999999999999999E-6,2.5000000000000002E-6,4.5035000000000002E-6,2.0281512250000002E-11,2.0000000000000002E-11,1.9864613600245953E-6]
[4,1.9999999999999999E-6,1.2500000000000001E-6,3.2535000000000003E-6,1.0585262250000003E-11,1.0000000000000001E-11,1.2442378269910875E-6]
[5,1.9999999999999999E-6,6.2500000000000005E-7,2.6284999999999999E-6,6.9090122499999995E-12,5.0000000000000005E-12,6.2341495275644866E-7]
[6,1.9999999999999999E-6,3.1250000000000003E-7,2.3159999999999999E-6,5.3638559999999994E-12,2.5000000000000003E-12,3.1185344059081598E-7]
[7,1.9999999999999999E-6,1.5625000000000001E-7,2.1597499999999999E-6,4.6645200624999996E-12,1.2500000000000001E-12,1.5595399925101828E-7]
[8,1.9999999999999999E-6,7.8125000000000006E-8,2.0816250000000002E-6,4.3331626406250003E-12,6.2500000000000006E-13,7.7982992685101703E-8]
[9,1.9999999999999999E-6,3.9062500000000003E-8,2.0425625000000001E-6,4.1720615664062506E-12,3.1250000000000003E-13,3.8992905570747062E-8]
[10,1.9999999999999999E-6,1.9531250000000002E-8,2.02303125E-6,4.0926554384765625E-12,1.5625000000000002E-13,1.949679472552353E-8]
[]
[]
[null,"When [Ligand] =< KD"]
[]
[null,"Req = (Rmax / (1 + (KD/C)))"]
[]
[null,"Req = value at equilibrium"]
[null,"Rmax = maximal value at equilibrium"]
[null,"KD = equilibrium dissociation constant",null,null,null,null,null,null,"1) Add equilibrium dissocation constant\n2) Add dilution step for variable component\n3) Add concentration constant component\n4) Add highest concentration variable component"]
[null,"C = concentration analyte"]
[]
[]
[null,"KD",1.1999999999999999E-7," (M-1)","Ligand",0.12," µM"]
[null,"Dilution",2," (x)","Analyte",10," µM"]
[]
[null,"Ligand","Analyte","Complex"]
[null,"FXI","FIX",{"$f":"CONCATENATE(B53,\"_\",C53)","v":"FXI_FIX"},"Sat.",{"$f":"CONCATENATE(B53,\":\",C53)","v":"FXI:FIX"}]
["n","(µM)","(µM)","(µM)","(%)","(1:x)"]
[1,{"$f":"F49","v":0.12},{"$f":"F50","v":10},{"$f":"(B55/1000000)/(1+(C$49/(C55/1000000)))*1000000","v":0.11857707509881421},{"$f":"(D55/B55)*100","v":98.814229249011845},{"$f":"C55/B55","v":83.333333333333343}]
[2,{"$f":"B55","v":0.12},{"$f":"C55/C$50","v":5},{"$f":"(B56/1000000)/(1+(C$49/(C56/1000000)))*1000000","v":0.11718749999999999},{"$f":"(D56/B56)*100","v":97.656249999999986},{"$f":"C56/B56","v":41.666666666666671}]
[3,{"$f":"B56","v":0.12},{"$f":"C56/C$50","v":2.5},0.11450381679389311,95.419847328244273,20.833333333333336]
[4,0.12,1.25,0.10948905109489049,91.240875912408754,10.416666666666668]
[5,0.12,0.625,0.10067114093959731,83.892617449664428,5.2083333333333339]
[6,0.12,0.3125,8.6705202312138727E-2,72.25433526011561,2.604166666666667]
[7,0.12,0.15625,6.7873303167420809E-2,56.561085972850677,1.3020833333333335]
[8,0.12,7.8125E-2,4.7318611987381708E-2,39.432176656151427,0.65104166666666674]
[9,0.12,3.90625E-2,2.9469548133595286E-2,24.557956777996072,0.32552083333333337]
[10,0.12,1.953125E-2,1.6797312430011199E-2,13.997760358342665,0.16276041666666669]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[]
[null,"Req = (Rmax / (1 + (KD/C)))"]
[]
[null,"Req ","complex concentratie"]
[null,"Rmax","ligand concentratie"]
[null,"C","analyte concentratie"]
[null,"KD","equilibrium dissociation constant"]
[]
[]
[null,"KD",1.1999999999999999E-7,"(M-1)","Ligand",1.9999999999999999E-6," M"]
[null,"Dilution",3,"(x)","Analyte",1.0000000000000001E-5," M"]
[]
[null,"Rmax","C","Req","Req","KD"]
["n","(M)","(M)","(M)","(%)","(x)"]
[1,{"$f":"F86","v":1.9999999999999999E-6},{"$f":"F87","v":1.0000000000000001E-5},{"$f":"(B91/(1+(C$86/C91)))","v":1.9762845849802369E-6},{"$f":"(D91/B91)*100","v":98.814229249011859},{"$f":"(C91/C$86)","v":83.333333333333343}]
[2,{"$f":"B91","v":1.9999999999999999E-6},{"$f":"C91/C$87","v":3.3333333333333337E-6},1.9305019305019305E-6,{"$f":"(D92/B92)*100","v":96.525096525096529},27.777777777777782]
[3,{"$f":"B92","v":1.9999999999999999E-6},1.1111111111111112E-6,1.8050541516245488E-6,90.25270758122744,9.2592592592592613]
[4,1.9999999999999999E-6,3.7037037037037042E-7,1.5105740181268883E-6,75.528700906344412,3.0864197530864206]
[5,1.9999999999999999E-6,1.234567901234568E-7,1.0141987829614603E-6,50.709939148073012,1.0288065843621401]
[6,1.9999999999999999E-6,4.1152263374485599E-8,5.1072522982635346E-7,25.536261491317674,0.34293552812071337]
[7,1.9999999999999999E-6,1.3717421124828532E-8,2.0517029134181371E-7,10.258514567090685,0.11431184270690445]
[8,1.9999999999999999E-6,4.5724737082761777E-9,7.3410659227719873E-8,3.6705329613859936,3.8103947568968149E-2]
[9,1.9999999999999999E-6,1.5241579027587259E-9,2.5084031505543576E-8,1.2542015752771787,1.2701315856322717E-2]
[10,1.9999999999999999E-6,5.0805263425290864E-10,8.4318453936828624E-9,0.42159226968414315,4.2337719521075724E-3]

{"sheet":"Solvent calibration","index":8,"n_rows":29,"n_cols":16,"rows_kept":29,"cut":false,"hidden":false,"merged":[[26,2,33,6],[4,2,4,3],[28,10,33,16],[11,6,11,8],[11,2,11,5],[5,2,5,3],[6,2,6,3],[7,2,7,3],[8,2,8,3],[9,2,9,3]]}
["BiaCalculations"]
["Solvent calculator"]
[null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"This page is not protected."]
[]
[null,null,"Solvent  "]
[null,null,"Solvent stock  ",null,10,"  (%)"]
[null,null,"no. Dilutions  ",null,6]
[null,null,"Lowest  ",null,2,"  (%)"]
[null,null,"Highest  ",null,6,"  (%)"]
[null,null,"Volume  ",null,500,"  (µl)"]
[]
[null,null,"Solvent calibration solutions",null,null,null,"Calibration response levels"]
[null,null,"Solvent","Buffer","Volume","Conc. ","Reference","Active","Correction"]
[null,null,"(µl)","(µl)","(µl)","(%)","(RU)","(RU)","(RU)"]
[null,1,{"$f":"E$10*E8/E$6","v":100},{"$f":"E$10-C15","v":400},{"$f":"C15+D15","v":500},{"$f":"C15*E$6/E15","v":2},-515,-487,{"$f":"H15-G15","v":28}]
[null,2,{"$f":"(E$10*((E$9-E$8)/(E$7-1))/E$6)*(B16-1)+C$15","v":140},360,{"$f":"C16+D16","v":500},2.8,-285,-265,{"$f":"H16-G16","v":20}]
[null,3,180,320,{"$f":"C17+D17","v":500},3.6,-85,-77,8]
[null,4,220,280,{"$f":"C18+D18","v":500},4.4000000000000004,171,169,-2]
[null,5,260,240,500,5.2,442,428,-14]
[null,6,300,200,500,6,642,617,-25]
[null,7,340,160,500,6.8,908,870,-38]
[null,8,380,120,500,7.6,1085,1038,-47]
[null,9,420,80,{"$f":"C23+D23","v":500},8.4,null,null,0]
[null,10,460,40,500,9.1999999999999993,null,null,0]
[]
[]
[null,null,"1) Set solvent stock concentration\n2) Set number of dilutions\n3) Set highest solvent control concentration\n4) Set lowest solvent control concentration\n5) Set volume of sample\n6) Add calibration response levels after measuring\n7) Use the correction values to correct measurements"]
[]
[null,null,null,null,null,null,null,null,"ref:",null,"Frostell-Karlsson, A., et al. (2000). \"Biosensor analysis of the interaction between immobilized human serum albumin and drug compounds for prediction of human serum albumin binding levels.\" J.Med.Chem. 43(10): 1986-1992."]

{"sheet":"Simulation","index":9,"n_rows":227,"n_cols":21,"rows_kept":227,"cut":false,"hidden":false,"merged":[[8,8,19,13]]}
["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Kinetic simulations / Association",null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Kinetic simulations / Dissociation"]
[]
[null,"Kinetic parameters",null,null,null,null,null,null,"Association formulas used",null,null,null,null,null,null,null,"Dissociation formula used"]
[null,null,null,null,null,null,null,null,"Rt = Req ( 1- e^ -(ka * C + kd)(t-t0)"]
[null,null,"Rt1","Rt2","Rt3","Rt4",null,null,"Req = (ka*C/ka*C+kd)*Rmax",null,null,null,null,null,null,null,"Rt=R0 * e-kd(t-t0)"]
[null,"Rmax",100,100,100,100]
[null,"ka",100000,100000,100000,100000,"M-1s-1"]
[null,"C",1.2499999999999999E-8,2.4999999999999999E-8,4.9999999999999998E-8,9.9999999999999995E-8,"M",null,"1) Add Rmax for the curves\n2) Add ka, analyte C and kd\n3) Add time step size (tstep) in seconds\n\n4) Req is calculated\n5) kobs (ka*C+kd, KD and C/KD are calculated\n\n6) tmax is calculated based on t step\n7) response at tmax and %response to Req and Rmax are calculated"]
[null,"kd",0.01,0.01,0.01,0.01,"s-1"]
[null,"Req",{"$f":"(C8*C9/(C8*C9+C10))*C7","v":11.111111111111112},{"$f":"(D8*D9/(D8*D9+D10))*D7","v":20},{"$f":"(E8*E9/(E8*E9+E10))*E7","v":33.333333333333336},{"$f":"(F8*F9/(F8*F9+F10))*F7","v":50}]
[null,"ka*C+kd",{"$f":"C8*C9+C10","v":1.125E-2},{"$f":"D8*D9+D10","v":1.2500000000000001E-2},{"$f":"E8*E9+E10","v":1.4999999999999999E-2},{"$f":"F8*F9+F10","v":0.02}]
[null,"KD",{"$f":"C10/C8","v":9.9999999999999995E-8},{"$f":"D10/D8","v":9.9999999999999995E-8},{"$f":"E10/E8","v":9.9999999999999995E-8},{"$f":"F10/F8","v":9.9999999999999995E-8},"M"]
[null,"C (x KD)",{"$f":"C9/C13","v":0.125},{"$f":"D9/D13","v":0.25},{"$f":"E9/E13","v":0.5},{"$f":"F9/F13","v":1},"x"]
[]
[null,"tmax",{"$f":"$B225","v":400},{"$f":"$B225","v":400},{"$f":"$B225","v":400},{"$f":"$B225","v":400},"s"]
[null,"R at tmax",{"$f":"C225","v":10.987677816241755},{"$f":"D225","v":19.865241060018292},{"$f":"E225","v":33.250708260777792},{"$f":"F225","v":49.983226868604874},"RU"]
[null,"% of Req",{"$f":"(C17/C11)*100","v":98.88910034617578},{"$f":"(D17/D11)*100","v":99.326205300091459},{"$f":"(E17/E11)*100","v":99.752124782333368},{"$f":"(F17/F11)*100","v":99.966453737209747},"%"]
[null,"% of Rmax",{"$f":"(C17/C7)*100","v":10.987677816241755},{"$f":"(D17/D7)*100","v":19.865241060018292},{"$f":"(E17/E7)*100","v":33.250708260777792},{"$f":"(F17/F7)*100","v":49.983226868604874},"%"]
[]
[null,"tstep"]
[null,2,"s"]
[]
[null,"t-to","Rt1","Rt2","Rt3","Rt4",null,null,null,null,null,null,null,null,null,null,"kd",{"$f":"C10","v":0.01},{"$f":"D10","v":0.01},{"$f":"E10","v":0.01},{"$f":"F10","v":0.01}]
[null,0,{"$f":"C$11*(1-(EXP(-((C$12*B25)))))","v":0},{"$f":"D$11*(1-(EXP(-((D$12*$B25)))))","v":0},0,0]
[null,{"$f":"B25+B$22","v":2},0.24720847562959622,0.4938017594333477,0.98514888171639492,1.9605280423838412,null,null,null,null,null,null,null,null,null,null,"t-to",{"$f":"C24","v":"Rt1"},{"$f":"D24","v":"Rt2"},{"$f":"E24","v":"Rt3"},{"$f":"F24","v":"Rt4"}]
[null,4,0.4889168685211116,0.97541150998571968,1.9411822138583761,3.8441826806682124,null,null,null,null,null,null,null,null,null,null,{"$f":"B25","v":0},{"$f":"C$225*EXP(-C$10*Q27)","v":10.987677816241755},{"$f":"D$225*EXP(-D$10*Q27)","v":19.865241060018292},{"$f":"E$225*EXP(-E$10*Q27)","v":33.250708260777792},{"$f":"F$225*EXP(-F$10*Q27)","v":49.983226868604874}]
[null,6,0.72524754871080577,1.4451302734289428,2.8689604909590605,5.6539781641421261,null,null,null,null,null,null,null,null,null,null,2,10.770107218202234,19.471882931948809,32.592300123724357,48.993492664197056]
[null,8,0.95632016365302031,1.9032516392808096,3.7693187760947509,7.3928105516894327,null,null,null,null,null,null,null,null,null,null,4,10.556844806653338,19.086313816680676,31.946929341290215,48.023356510110204]
[null,10,1.1822516987942704,2.3500619483080909,4.6430674524980731,9.0634623461009092,null,null,null,null,null,null,null,null,null,null,6,10.347805273786912,18.708379481426793,31.314337756557322,47.072430338947356]
[null,12,1.4031565367996177,2.7858404714988438,5.4909929529576003,10.668606946672327,null,null,null,null,null,null,null,null,null,null,8,10.142905001002577,18.337928747413866,30.694272324457124,46.140333767560854]
[null,14,1.6191465154613389,3.2108595846158527,6.313858467660431,12.210812927213727,null,null,null,null,null,null,null,null,null,null,10,9.9420620254591796,17.974813429408879,30.086485010549126,45.226693944894031]
[null,16,1.8303309843192002,3.6253849384403636,7.1124046311148845,13.692548146315453,null,null,null,null,null,null,null,null,null,null,12,9.7451960072885608,17.61888827644303,29.490732691803835,44.331145402835794]
[null,18,2.0368168600209908,4.0296756248124588,7.8873501887715625,15.116183696448449,null,null,null,null,null,null,null,null,null,null,14,9.5522281974585361,17.270010913709406,28.906777059350279,43.453329910028515]
[null,20,2.2387086804513663,4.423984338571902,8.6393926439427382,16.483997698218033,null,null,null,null,null,null,null,null,null,null,16,9.3630814062722365,16.928041785611143,28.334384523149289,42.592896328570696]
[null,22,2.4361086576563813,4.8085575355006309,9.3692088856024611,17.798178945842931,null,null,null,null,null,null,null,null,null,null,18,9.1776799724911946,16.592844099937359,27.773326118554394,41.749500473557163]
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["BiaCalculations",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"BiaCalculations"]
["Kinetic plot",null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,null,"Kinetic plot"]
[]
[null,null,"Nanobodies over FXI",null,null,null,null,null,null,null,"FXIIa over FXIwt"]
[]
[]
[null,null,"1A5",null,"1B3",null,"2E3",null,"2H5",null,"1H9",null,"2G3"]
[null,null,"ka","kd","ka","kd","ka","kd","ka","kd","ka","kd","ka","kd","ka","kd","ka","kd"]
["Experiment","fit-model","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)","(M-1s-1)","(s-1)"]
["1A5","Apple 3",34300,9.7799999999999992E-4,42700,5.04E-4,35400,7.9900000000000001E-4,73300,1.1400000000000001E-4,109000,1.08E-4,124000,9.8999999999999999E-4]
["1B3","FXI",87300,5.1800000000000001E-4,59400,8.3499999999999997E-5,87000,7.36E-4,262000,7.7000000000000001E-5,309000,7.8999999999999996E-5,425000,1.1069999999999999E-3]
[" 2E3","FXIa",86500,5.1699999999999999E-4,64100,7.7899999999999996E-5,88100,7.7999999999999999E-4,291000,7.2299999999999996E-5,335000,8.3300000000000005E-5,435000,1.09E-3]
["2H5"]
["1H9"]
["2G3"]
[]
[8]
[9]
[10]
[11]
[12]
[13]
[14]
[15]
[16]
[17]
[18]
[19]
[20]
[]
[]
[null,"Each kinetic pair will give a point."]
[null,"In this plot eight different groups are defined."]
[null,"Change values as you need."]