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Development of an antigen microarray for high throughput monoclonal antibody selection.

Abstract：

:Monoclonal antibodies are valuable laboratory reagents and are increasingly being exploited as therapeutics to treat a range of diseases. Selecting new monoclonal antibodies that are validated to work in particular applications, despite the availability of several different techniques, can be resource intensive with uncertain outcomes. To address this, we have developed an approach that enables early screening of hybridoma supernatants generated from an animal immunised with up to five different antigens followed by cloning of the antibody into a single expression plasmid. While this approach relieved the cellular cloning bottleneck and had the desirable ability to screen antibody function prior to cloning, the small volume of hybridoma supernatant available for screening limited the number of antigens for pooled immunisation. Here, we report the development of an antigen microarray that significantly reduces the volume of supernatant required for functional screening. This approach permits a significant increase in the number of antigens for parallel monoclonal antibody selection from a single animal. Finally, we show the successful use of a convenient small-scale transfection method to rapidly identify plasmids that encode functional cloned antibodies, addressing another bottleneck in this approach. In summary, we show that a hybrid approach of combining established hybridoma antibody technology with refined screening and antibody cloning methods can be used to select monoclonal antibodies of desired functional properties against many different antigens from a single immunised host.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Staudt N,Müller-Sienerth N,Wright GJ

doi

10.1016/j.bbrc.2013.12.033

subject

Has Abstract

pub_date

2014-03-21 00:00:00

pages

785-90

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(13)02091-3

journal_volume

445

pub_type

杂志文章

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