血清中免疫球蛋白G和Fab的荧光素缀合物的微量制备物。

A micropreparation of fluorescein conjugates of immunoglobulin G and Fab from serum.

作者信息

Mohanty J G, Rosenthal K S

出版信息

Anal Biochem. 1985 May 1;146(2):361-5. doi: 10.1016/0003-2697(85)90552-4.

Abstract

Protein A-Sepharose CL-4B (PAS) was used to isolate rabbit immunoglobulin G from crude anti-herpes simplex virus-1 serum. Papain treatment of the PAS-bound immunoglobulin G released Fab fragments from the solid support, while Fc-containing fragments remained bound to PAS. PAS-immobilized immunoglobulin G was fluoresceinated by reaction with fluorescein isothiocyanate followed by papain cleavage to yield fluorescein-conjugated Fab fragments in solution. These fragments retained activity toward herpes simplex virus-1 infected Vero cells as evaluated by immunofluorescence. This novel procedure represents the fastest and simplest method for preparing Fab or fluoresceinated Fab fragments directly from any volume of immune serum.

摘要

蛋白A-琼脂糖凝胶CL-4B（PAS）用于从抗单纯疱疹病毒-1粗血清中分离兔免疫球蛋白G。对结合在PAS上的免疫球蛋白G进行木瓜蛋白酶处理，可从固相支持物上释放出Fab片段，而含Fc的片段仍结合在PAS上。将结合在PAS上的免疫球蛋白G与异硫氰酸荧光素反应进行荧光素化，然后用木瓜蛋白酶裂解，在溶液中产生荧光素偶联的Fab片段。通过免疫荧光评估，这些片段对单纯疱疹病毒-1感染的Vero细胞仍具有活性。这种新方法是直接从任何体积的免疫血清中制备Fab或荧光素化Fab片段的最快、最简单的方法。