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A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells. / Guselnikov, Sergey V.; Belovezhets, Tatyana N.; Kulemzin, Sergey V. et al.

In: BioTechniques, Vol. 67, No. 4, 01.10.2019, p. 184-187.

Research output: Contribution to journal › Article › peer-review

Harvard

Guselnikov, SV, Belovezhets, TN, Kulemzin, SV, Gorchakov, AA & Taranin, AV 2019, 'A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells', BioTechniques, vol. 67, no. 4, pp. 184-187. https://doi.org/10.2144/btn-2019-0079

APA

Guselnikov, S. V., Belovezhets, T. N., Kulemzin, S. V., Gorchakov, A. A., & Taranin, A. V. (2019). A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells. BioTechniques, 67(4), 184-187. https://doi.org/10.2144/btn-2019-0079

Vancouver

Guselnikov SV, Belovezhets TN, Kulemzin SV, Gorchakov AA, Taranin AV. A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells. BioTechniques. 2019 Oct 1;67(4):184-187. doi: 10.2144/btn-2019-0079

Author

Guselnikov, Sergey V. ; Belovezhets, Tatyana N. ; Kulemzin, Sergey V. et al. / A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells. In: BioTechniques. 2019 ; Vol. 67, No. 4. pp. 184-187.

BibTeX

@article{5ec58f4d51484c619172ecc8f430173a,
title = "A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells",
abstract = "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.",
keywords = "B lymphocytes, heavy chain, immunoglobulin, light chain, single-cell PCR",
author = "Guselnikov, {Sergey V.} and Belovezhets, {Tatyana N.} and Kulemzin, {Sergey V.} and Gorchakov, {Andrey A.} and Taranin, {Alexander V.}",
year = "2019",
month = oct,
day = "1",
doi = "10.2144/btn-2019-0079",
language = "English",
volume = "67",
pages = "184--187",
journal = "BioTechniques",
issn = "0736-6205",
publisher = "Eaton Publishing Company",
number = "4",

}

RIS

TY - JOUR

T1 - A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells

AU - Guselnikov, Sergey V.

AU - Belovezhets, Tatyana N.

AU - Kulemzin, Sergey V.

AU - Gorchakov, Andrey A.

AU - Taranin, Alexander V.

PY - 2019/10/1

Y1 - 2019/10/1

N2 - 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.

AB - 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.

KW - B lymphocytes

KW - heavy chain

KW - immunoglobulin

KW - light chain

KW - single-cell PCR

UR - http://www.scopus.com/inward/record.url?scp=85072716932&partnerID=8YFLogxK

U2 - 10.2144/btn-2019-0079

DO - 10.2144/btn-2019-0079

M3 - Article

C2 - 31411041

AN - SCOPUS:85072716932

VL - 67

SP - 184

EP - 187

JO - BioTechniques

JF - BioTechniques

SN - 0736-6205

IS - 4

ER -

ID: 21699879