A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells

Sergey V. Guselnikov, Tatyana N. Belovezhets, Sergey V. Kulemzin, Andrey A. Gorchakov, Alexander V. Taranin

Результат исследования: Научные публикации в периодических изданияхстатья

Аннотация

Cloning VH and VL genes from individual antigen-specific B cells is an attractive approach for producing monoclonal antibodies of the desired specificity. Current RT-PCR protocols, however, result in the successful identification of VH and VL gene pairs in about half of the sorted cells. Here, we demonstrate that single-cell RT-PCR is likely affected by stochastic factors, and that running PCRs in triplicate results in successful amplification of the expressed VH and VL genes in 90-100% of single sorted human B cells.

Язык оригиналаанглийский
Страницы (с-по)184-187
Число страниц4
ЖурналBioTechniques
Том67
Номер выпуска4
DOI
СостояниеОпубликовано - 1 окт 2019

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