Analysis of kinetic data of antibody-antigen interaction from an optical biosensor by exponential curve fitting

被引:20
作者
Gill, A
Leatherbarrow, RJ
Hoare, M
PollardKnight, DV
Lowe, PA
Fortune, DH
机构
[1] FISONS APPL SENSOR TECHNOL,CAMBRIDGE CB3 8SL,ENGLAND
[2] UNIV LONDON IMPERIAL COLL SCI TECHNOL & MED,DEPT CHEM,LONDON SW7 2AY,ENGLAND
基金
英国生物技术与生命科学研究理事会;
关键词
optical biosensor; kinetic analysis; exponential curve fitting; biomolecular interaction; recombinant antibody fragments;
D O I
10.1016/0168-1656(96)01410-1
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An optical biosensor system employing a resonant mirror (RM), with a stirred cuvette has been used to follow the interaction of a recombinant antibody fragment with its antigen, hen egg lysozyme. The data generated by the biosensor were analysed in order to determine the kinetic constants for the interaction using a linear transform (derivative analysis). For comparison the data were also analysed using an exponential curve fitting routine. It was demonstrated that the exponential curve fitting method produced results which were in agreement with the existing linear transform method. It was also shown that early fitting of the association phase response, using the exponential curve fitting routine between 0 and 70 s after sample addition, yielded sufficient information to provide a prediction of k(on). The potential use of the optical biosensor for the rapid monitoring of protein production and purification is discussed.
引用
收藏
页码:117 / 127
页数:11
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