Covalent immobilization of antibodies for the preparation of immunoaffinity chromatographic supports

被引：4

作者：

Alberto Mejia-Manzano, Luis ^{[1
]}

Gonzalez-Valdez, Jose ^{[1
]}

Mayolo-Deloisa, Karla ^{[1
]}

Escalante-Vazquez, Edgardo J. ^{[2
]}

Rito-Palomares, Marco ^{[1
]}

机构：

[1] Tecnol Monterrey, Ctr Biotecnol FEMSA, Escuela Ingn & Ciencias, Campus Monterrey, Monterrey, NL, Mexico

[2] Tecnol Monterrey, Dept Ingn Ind, Campus Monterrey, Monterrey, NL, Mexico

来源：

SEPARATION SCIENCE AND TECHNOLOGY | 2016年 / 51卷 / 10期

关键词：

Coupling efficiency (CE); IgG; IgM; immunoaffinity chromatography (IAC); optimization; AFFINITY-CHROMATOGRAPHY; SOLID-PHASE; MONOCLONAL-ANTIBODIES;

D O I：

10.1080/01496395.2016.1174264

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Immunosorbents in immunoaffinity chromatography (IAC) are prepared by immobilizing expensive antibodies without guidelines for ensuring the best coupling efficiencies, and avoiding low binding capacities. Covalent immobilization of antibodies on N-hydroxysuccinimide (NHS)-activated Sepharose 4 Fast Flow resin was optimized using human IgG via full factorial design with incubation times (4, 9, 14, 19 and 24 h), temperatures (4 degrees C and 20 degrees C) and coupling reaction buffers (sodium bicarbonate and triethanolamine). The best coupling efficiency (CE) (83.4 +/- 8.7%) was reached with triethanolamine buffer, 14 h and 4 degrees C. Comparison of antibody isotypes (IgG or IgM) by a nested factorial analysis suggested that antibodies in the IgG isotype presents the best coupling efficiency.

引用

页码：1736 / 1743

页数：8

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