Technical Service

Hybridoma Sequencing

Introduction

Hybridoma antibody technology refers to the preparation of monoclonal antibodies through the fusion of B lymphocytes and myeloma cells.

Hybridoma antibody technology is widely used and mature, but due to the instability of hybridomas in the process of transmission, which affects the production of antibodies; hybridoma cells require high preservation conditions and need to be frozen in liquid nitrogen for a long period of time; the monoclonal antibody sequence produced by hybridomas can be obtained through hybridoma sequencing, which can be used for the production of antibodies by means of recombinant expression, which solves the problem of instability and difficulty in preservation of hybridoma cells, and produces antibodies by means of in vitro recombinant expression, which ensures the stability of antibodies, and at the same time, effectively produces antibodies by means of in vitro recombinant expression, which effectively ensures the stability of antibodies. This solves the problem of instability and difficulty in preservation of hybridoma cells, and produces antibodies by recombinant expression in vitro, which ensures the stability of antibodies and effectively avoids endogenous biological contamination.

AtaGenix provides an overall solution from hybridoma sequencing to recombinant antibody expression: high sequencing accuracy, 2 weeks turnaround time. We could reach up to 200-500mg/L in transient expression by using the third-generation proprietary vector pATX3.0 and mammalian cells.

Advantages

1. Accepts cells in culture as well as frozen cell lines

2. Short cycle time: only 2 weeks

3. Variable region sequencing and full-length sequencing are available.

4. Experiment report (including CDR region analysis).

Contents

Projects

Receivables

Contents

Time Frame

Deliverables

Antibody variable

region/

full-length sequencing

1x106Hybridoma cells

 

Monoclonal antibody subtype

RNA extraction

mRNA reverse transcription c DNA

PCR amplification of antibody heavy and light chain genes

 

Cloning and sequencing of heavy and light chain genes

 

Bioinformatics analysis of

sequencing results to determine functional antibody genes and

antibody CDR regions

 

Experimental report

    2-3 Weeks   

Sequencing report

 

Sequence original data

 

Heavy/Light chain analysis

 

Plasmids containing antibody fragments