Amplification of IgG VH and VL (Fab) from single human plasma cells and B cells

被引：36

作者：

Coronella, J. A.

Telleman, P.

Truong, T. D.

Ylera, F.

Junghans, R. P. ^{[1
]}

机构：

[1] Beth Israel Deaconess Med Ctr, Boston, MA 02215 USA

来源：

NUCLEIC ACIDS RESEARCH | 2000年 / 28卷 / 20期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1093/nar/28.20.e85

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Amplification of human immunoglobulin has many potential applications such as analysis of clonality, isolation of immunogenic antigens and antigen-specific immunotherapy. Here we describe a method for amplification of human immunoglobulin heavy and light chains from single B lymphocytes or plasma cells. Cells are isolated by FACS, and Ig is amplified by semi-nested RT-PCR. The method is versatile, sensitive and reliable: it provides appropriately paired heavy and light chains, requiring as little as 2 days to produce amplified Fab DNA from human tissues.

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页数：7

相关论文

共 13 条

[1] A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities [J].

Babcook, JS ;

Leslie, KB ;

Olsen, OA ;

Salmon, RA ;

Schrader, JW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (15) :7843-7848

[2] HUMAN-ANTIBODIES FROM COMBINATORIAL LIBRARIES [J].

BURTON, DR ;

BARBAS, CF .

ADVANCES IN IMMUNOLOGY, VOL 57, 1994, 57 :191-280

[3] A new method for the analysis and production of monoclonal antibody fragments originating from single human B cells [J].

deWildt, RMT ;

Steenbakkers, PG ;

Pennings, AHM ;

vandenHoogen, FHJ ;

vanVenrooij, WJ ;

Hoet, RMA .

JOURNAL OF IMMUNOLOGICAL METHODS, 1997, 207 (01) :61-67

[4] ANTIBODY REDESIGN BY CHAIN SHUFFLING FROM RANDOM COMBINATORIAL IMMUNOGLOBULIN LIBRARIES [J].

KANG, AS ;

JONES, TM ;

BURTON, DR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (24) :11120-11123

[5]

Kantor AB, 1995, ANN NY ACAD SCI, V764, P224

[6] Single-cell analysis by RT-PCR reveals differential expression of multiple type 1 and 2 cytokine genes among cells within polarized CD4+ T cell populations [J].

Kelso, A ;

Groves, P ;

Ramm, L ;

Doyle, AG .

INTERNATIONAL IMMUNOLOGY, 1999, 11 (04) :617-621

[7]

LAGERKVIST ACS, 1995, BIOTECHNIQUES, V18, P862

[8] SEQUENCING HEAVY-CHAIN AND LIGHT-CHAIN VARIABLE GENES OF SINGLE B-HYBRIDOMA CELLS BY TOTAL ENZYMATIC AMPLIFICATION [J].

LIU, AH ;

CREADON, G ;

WYSOCKI, LJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (16) :7610-7614

[9] Phage libraries - A new route to clinically useful antibodies [J].

Marks, C ;

Marks, JD .

NEW ENGLAND JOURNAL OF MEDICINE, 1996, 335 (10) :730-733

[10] Bcl-2(+) tonsillar plasma cells are rescued from apoptosis by bone marrow fibroblasts [J].

Merville, P ;

Dechanet, J ;

Desmouliere, A ;

Durand, I ;

deBouteiller, O ;

Garrone, P ;

Banchereau, J ;

Liu, YJ .

JOURNAL OF EXPERIMENTAL MEDICINE, 1996, 183 (01) :227-236