A SIMPLE STANDARDIZED PROTOCOL FOR THE PRODUCTION OF MONOCLONAL-ANTIBODIES AGAINST VIRAL AND BACTERIAL-ANTIGENS

被引:13
作者
COYLE, PV
WYATT, D
MCCAUGHEY, C
ONEILL, HJ
机构
[1] Regional Virus Laboratory, Royal Victoria Hospital, Belfast
关键词
MONOCLONAL ANTIBODY; HYBRIDOMA TECHNIQUE; HUMAN RESPIRATORY VIRUS; HELICOBACTER-PYLORI;
D O I
10.1016/0022-1759(92)90308-G
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A simple standardised protocol for making monoclonal antibodies against a range of human bacteria and viruses is described. The protocol was designed to reduce the number of steps to a minimum. A one step footpad immunisation was followed by the fusion schedule 10-15 days later. A vital step in the technique was the use of the immunised mouse's spleen to provide a feeder layer post fusion. This simplified the protocol and more importantly greatly accelerated the growth of the hybridomas produced. Immunisation, fusion and clonal expansion of specific antibody secreting hybridomas was complete within 5 weeks. The percentage of hybridomas secreting specific antibody ranged from 6% to 28%, the majority of which were of the IgG isotypes. The method was economical in the use of tissue culture medium and simple to perform.
引用
收藏
页码:81 / 84
页数:4
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