Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells

被引：6
|
作者：
Viant, Charlotte [1 ]
Escolano, Amelia [1 ]
Chen, Spencer T. [1 ]
Nussenzweig, Michel C. [1 ,2 ]
机构：
[1] Rockefeller Univ, Lab Mol Immunol, New York, NY 10065 USA
[2] Rockefeller Univ, Howard Hughes Med Inst HHMI, New York, NY 10065 USA
来源：
STAR PROTOCOLS | 2021年 / 2卷 / 02期
关键词：
Protocol describes; BCR analysis; somatic; relationships; antigen; MEMORY;
D O I：
10.1016/j.xpro.2021.100389
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
The analysis of B cell receptors (BCR) from single B cells is crucial to understanding humoral immune responses. Here, we describe a protocol for the sequencing, cloning, and characterization of antibody genes that encode BCRs. We used this method to analyze the BCRs of different mouse B cell populations for somatic hypermutations, clonal and phylogenic relationships, and their affinity for cognate antigen. For complete details on the use and execution of this protocol, please refer to Viant et al. (2020).
引用
收藏
页数：23
相关论文
共 50 条
  • [31] MONOCLONAL-ANTIBODIES BCA-B/20 TO MEMBRANOUS ANTIGEN OF HUMAN B-CELLS
    BUROVA, GF
    MITEREV, GY
    KORSHUNOV, LI
    BULYCHEVA, TI
    GEMATOLOGIYA I TRANSFUZIOLOGIYA, 1990, 35 (08): : 28 - 29
  • [32] Efficient generation single human B cells by of monoclonal antibodies from single cell RT-PCR and expression vector cloning (vol 329, pg 112, 2008)
    Tiller, Thomas
    Meffre, Eric
    Yurasov, Sergey
    Tsuiji, Makoto
    Nussenzweig, Michel C.
    Wardemann, Hedda
    JOURNAL OF IMMUNOLOGICAL METHODS, 2008, 334 (1-2) : 142 - 142
  • [33] Properties and function of polyreactive antibodies and polyreactive antigen-binding B cells
    Zhou, Zhao-Hua
    Tzioufas, Athanasios G.
    Notkins, Abner Louis
    JOURNAL OF AUTOIMMUNITY, 2007, 29 (04) : 219 - 228
  • [34] ANALYSIS OF HUMAN AND MOUSE TATA-BINDING PROTEINS, USING MONOCLONAL-ANTIBODIES
    THOMPSON, N
    STRASHEIM, L
    NOLAN, K
    SLANSKY, J
    FARNHAM, P
    BURGESS, R
    JOURNAL OF CELLULAR BIOCHEMISTRY, 1993, : 94 - 94
  • [35] Generation of recombinant human monoclonal antibodies to rotavirus from single antigen-specific B cells selected with fluorescent virus-like particles
    Weitkamp, JH
    Kallewaard, N
    Kusuhara, K
    Feigelstock, D
    Feng, NG
    Greenberg, HB
    Crowe, JE
    JOURNAL OF IMMUNOLOGICAL METHODS, 2003, 275 (1-2) : 223 - 237
  • [36] Development and application of classical swine fever virus monoclonal antibodies derived from single B cells
    Zhongyuan Ma
    Yongcong Zhao
    Jianliang Lv
    Li Pan
    Veterinary Research, 54
  • [37] Development and application of classical swine fever virus monoclonal antibodies derived from single B cells
    Ma, Zhongyuan
    Zhao, Yongcong
    Lv, Jianliang
    Pan, Li
    VETERINARY RESEARCH, 2023, 54 (01) : 90
  • [38] THE STRUCTURE OF THE VARIABLE REGIONS OF MOUSE MONOCLONAL-ANTIBODIES TO HEPATITIS-B VIRUS CORE ANTIGEN
    SKRIVELIS, V
    STEINBERG, Y
    BICHKO, V
    GREN, E
    TSIMANIS, A
    SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 1993, 37 (06) : 637 - 643
  • [39] A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities
    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
  • [40] EFFICIENT GENERATION OF MONOCLONAL-ANTIBODIES FROM PRESELECTED ANTIGEN-SPECIFIC B-CELLS - EFFICIENT IMMORTALIZATION OF PRESELECTED B-CELLS
    STEENBAKKERS, PGA
    HUBERS, HAJM
    RIJNDERS, AWM
    MOLECULAR BIOLOGY REPORTS, 1994, 19 (02) : 125 - 134
12345