RT-PCR FACS Micromanipulators Single B Cell Antibody Cloning

Introduction

The immune system is a very robust “antibody factory” that consistently produces antibodies to fight infections and diseases by recognizing and targeting infected cells or pathogens (antigens). The emergence of single B cell antibody technology has enabled a new strategy for discovery of naturally occurring antibody drugs. The single B cell antibody technology produces higher throughput and higher coverage than the conventional hybridoma or phage display technologies. Most importantly, this technology also avoids the disadvantage of unnaturally paired heavy and light antibody chain displayed by phage.

With our advanced technology and professional team, we have developed two methods for discovering single B cell antibodies. The first method is based on the antigen-specific single B cells sorting by Fluorescence Activated Cell Sorter (FACS), followed by RT-PCR amplification to obtain the target antibody sequence. The second method relies on the use of micromanipulators to select a single B cell of interest, then culturing the cell for further characterization. With these two complementary methods and our other custom antibody discovery services, we help our customers effectively find antibody drugs while saving both time and money.

General procedure of single B cell antibody cloning and sequencing

* Extract and purify total RNAs from individual B cells;
* Synthesize cDNAs using optimized primer sets;
* Amplify DNA fragments including VH and VL, leader sequences, and partial constant regions (CH and CL) using 5' RACE;
* Sequence the amplified DNA fragments.

Service content

Required materials

Single or clonal B cells (contact technical services for details).

Deliverables

Study report including the number of antibody heavy and light chains identified, the DNA and protein sequences for each chain, and CDR analysis. Leader sequences and the amplified DNA fragments available if requested.

Service Specification

Key Features

• Ultra-high-throughput screening of B cells for antibody selection
• Unprecedented single-cell resolution
• Ideal for important and challenging target cells
• Available for most species