Antibody Variable Region Sequencing and Cloning

The sequencing of antibody variable regions (VH and VL) from hybridomas or clonal B cells is a critical first step in numerous applications, including enabling production of recombinant antibody in various formats, antibody engineering, humanization, and affinity maturation. LakePharma can sequence variable heavy and light chain regions from hybridoma cell lines or clonal primary B cells from a variety of species. Following this service, the variable regions can be reformatted into whole IgGs or single-chain (scFv)-Fc and transiently expressed in mammalian cells, followed by confirmation of antigen binding.

Please contact LakePharma at inquiries@lakepharma.com for custom quotes and details.

Cat.# Service Name Timeline Size Price Quantity Request
24101 Cloning and sequencing of mouse hybridoma IgG variable regions 1 week TBD $1500
24102 Cloning and sequencing of rat hybridoma IgG variable regions 1-2 weeks TBD $2400
24103 Cloning and sequencing of rabbit hybridoma IgG variable regions 1-2 weeks TBD $2400
24104 Cloning and sequencing of human hybridoma IgG variable regions 1-2 weeks TBD $2400
24105 Cloning and sequencing of primate B cell IgG variable regions 1-2 weeks TBD $2400
24106 Cloning and sequencing of hamster hybridoma IgG variable regions 1-2 weeks TBD $3000
24111 Cloning and sequencing of mouse hybridoma IgM variable regions 1-2 weeks TBD $3000
24112 Cloning and sequencing of rat hybridoma IgM variable regions 1-2 weeks TBD $3000
24114 Cloning and sequencing of primate B cell IgM variable regions 1-2 weeks TBD $3000

Cloning and sequencing antibody variable regions:

1. Extract and purify total RNA from hybridoma cells or clonal B cell.

2. Synthesize cDNA using optimized primer set.

3. Amplify the DNA fragments corresponding to the VL and VH using nested PCR.

4. Sequence the amplified DNA fragments. Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.

For cloning and sequencing antibody V regions:
A. Greater than 100,000 cells in frozen vial(s) and shipped on dry ice

OR

B. Isolated RNA samples dried on RNA isolation columns (before elution) and shipped on dry ice.

 

Note: please see FAQ: "For antibody variable region cloning, how should I prepare my hybridoma/B cell sample?"

Please include a hybridoma sample submission form.

1. Study report containing sequence analysis

2. Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.