THE MEASUREMENT OF RELATIVE ANTIBODY-AFFINITY BY ELISA USING THIOCYANATE ELUTION
THE MEASUREMENT OF RELATIVE ANTIBODY-AFFINITY BY ELISA USING THIOCYANATE ELUTION
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DOI：
10.1016/0022-1759(88)90196-2
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发表时间：
1988-02-10
影响因子：
2.2
通讯作者：
JONES, CL
中科院分区：
文献类型：
--
作者：
MACDONALD, RA;HOSKING, CS;JONES, CL
A variation of the standard ELISA assay was used to determine the relative affinities of six murine monoclonal anti-dinitrophenol (DNP) antibodies for DNP-bovine serum albumin (DNP-BSA). The procedure involved exposing replicate wells containing antibody bound to antigen to increasing concenrations of the chaotropic thiocyanate ion. Resistance to thiocyanate elution was utilized as a measure of affinity and an index (affinity index) representing a 50% reduction in initial absorbance was used to rank the anti-DNP antibodies in order of increasing affinity. For comparison, the affinity constants of the six monoclonals were determined by equilibrium dialysis with 3H-.epsilon.-DNP-L-lysine as antigen. A significant correlation (P < 0.02) was observed between the ranking orders obtained using the two methods, thereby demonstrating the applicability of the elution technique for the measurement of relative antibody affinity.
