Sequencing and cloning of antigen-specific antibodies from mouse …
Here we describe a protocol to sequence and clone the variable antibody region of single antigen-specific mouse memory B cells for antibody production. After purification, antigen-specific mouse memory B cells are first single-cell-sorted by fluorescence-activated cell sorting (FACS), and V (D)J transcripts are amplified by RT-PCR.
A simplified workflow for monoclonal antibody sequencing
2019年6月24日 · We successfully sequenced the variable regions of five mouse monoclonal IgG antibodies using this method, which enabled us to design chimeric mouse/human antibody expression plasmids for recombinant antibody production in …
Mass Spectrometry-Based De Novo Sequencing of Monoclonal Antibodies …
2021年6月14日 · We applied the method to sequence the widely used anti-FLAG-M2 mouse monoclonal antibody, which we successfully validated by remodeling a high-resolution crystal structure of the Fab and demonstrating binding to a FLAG-tagged target protein in Western blot analysis. The method thus offers robust and reliable sequences of monoclonal antibodies.
Sequencing and cloning of antigen-specific antibodies from mouse memory ...
2016年9月15日 · Here we describe a protocol to sequence and clone the variable antibody region of single antigen-specific mouse memory B cells for antibody production. After purification, antigen-specific...
Sequencing, cloning, and antigen binding analysis of monoclonal ...
2021年6月6日 · 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.
Hybridoma antibody cloning and sequencing protocols
2016年5月23日 · The first method established to use PCR to amplify and clone mouse immunoglobulin (Ig) variable (V) regions after selective first-strand cDNA synthesis. The procedure of hybridoma antibody cloning and sequencing sounds very straightforward. However, many factors affects the efficiency and success rate.
Efficient amplification and direct sequencing of mouse variable …
1999年6月11日 · This strategy could be a powerful tool for antibody sequence assessment whatever the V gene family before humanization of mouse monoclonal antibody or identification of paratope-derived peptides.
Large-scale network analysis reveals the sequence space ... - Nature
2019年3月21日 · Although antibody sequence diversity varied highly among mice (74–85% unique clones in a given mouse, Supplementary Fig. 2a), we found a remarkable cross-mouse consistency in clonal ...
Mouse Antibody - labome.com
2022年11月20日 · The mouse is the predominant animal host for monoclonal antibody production and also a host for polyclonal antibody generation. While typical mouse monoclonal antibody production involves immunizing mice (often BALB/c mice, for example [289, 290] ) with foreign antigens along with the commonly used Freund adjuvant or the new, water-soluble ...
NAb-seq: an accurate, rapid, and cost-effective method for antibody ...
2022年8月13日 · Here, we leveraged recent improvements in long-read Oxford Nanopore Technologies (ONT) sequencing to develop Nanopore Antibody sequencing (NAb-seq): a three-day, species-independent, and cost-effective workflow to characterize paired full-length immunoglobulin light- and heavy-chain genes from hybridoma cell lines.