测定手册指南_Assay_Handbook.pdf

GEHealthcare

LifeSciences

Biacore™AssayHandbook

1Introduction

1.1WhatBiacore™systemsmeasure5

1.2WhereBiacoresystemsareused5

1.3Scopeofthisbook6

1.4Biacoresystemrange6

1.5Biacoreterminology7

2Applicationoverview

2.1Screeninganddetectingbindingpartners9

2.2Kineticsandaffinitymeasurements10

2.2.1Kineticanalysis10

2.2.2Affinityanalysis11

2.3Concentrationmeasurements13

2.3.1Calibratedassays13

2.3.2Calibration-freeconcentrationanalysis(CFCA)14

2.4Mappingbindingsites14

3Generalconsiderations

3.1Sensorsurfaces17

3.1.1Generalproperties17

3.1.2Sensorsurfacetypes17

3.2Attachingtheligand19

3.2.1Covalentimmobilization19

3.2.2Highaffinitycapture21

3.2.3Responselevels21

3.3Generalbufferconsiderations23

3.3.1Buffersubstances23

3.3.2Ionicstrength23

3.3.3Additives24

3.3.4Bufferpreparation25

3.4Matchingsampleandrunningbuffer26

3.4.1Matchingrefractiveindex26

3.4.2Matchingbuffercomposition27

3.4.3Matchingbufferenvironmentinpractice28

3.5Dilutionseriesandreplicates28

3.5.1Dilutionseries28

3.5.2Replicates29

3.6Preparingvialsandmicroplate