Hybridoma Sequencing Service

The acquisition of the monoclonal antibody gene sequence of hybridoma cells is the basis for the preparation of recombinant antibodies and the development of antibody drugs. KMD Bioscience provides accurate and fast monoclonal antibody sequencing services to ensure that the antibody of interest can be accurately reconstituted before it enters downstream applications.

In the process of preparing recombinant antibodies, after the hybridoma cells are supplied by the customer, there are two heavy chain and two light chain problems, which leads to the diversity of subsequent light and heavy chain pairing combinations, making the preparation of recombinant antibodies time-consuming and labor-intensive. . Our scientists have proposed a very good solution. We designed a series of IgG primers for myeloma cells, combined with cell subcloning technology to finally obtain sequence information that is highly consistent with the target antibody.

 

Service Contents:

--- Standard monoclonal antibody sequencing services: antibody variable region VH and VL sequencing, antibody VH, VL and leader sequencing, whole antibody VH, VL, leader and constant region sequencing, sequencing services completed in 3-4 weeks.

--- Rapid monoclonal antibody sequencing service: The antibody sequence rapidly recognized from the target cell line can be obtained in 10 days.

 

Hybridoma Sequencing Service Procedure:

 

Step

Contents

Timeline

1

Hybridoma cells expended culture and antibody titer tested by ELISA

1-2 weeks

2

Total RNA extracted after Trizol cleavage

1-2 days

3

Reverse transcription preparation of cDNA by RT-PCR

1-2 days

4

The full-length gene [VHCH123 and VKCK/VLCL] genes were amplified by PCR to ensure the sequence fidelity of the variable regions FR1 and FR4.

1week

5

The T vector was sequenced, analyze the sequencing results and save the right clone.

3-5 days

 

Service Highlights:

*Using high-fidelity polymerase to obtain all sequence information of monoclonal antibody genes

*Personalized monoclonal antibody sequencing

*Multi-species hybridoma sequencing: mice, rats, rabbits, etc.

*Complete and rigorous bioinformatics analysis

*Customer has complete intellectual property and information security (high confidentiality)

*IgG whole gene cloning

 

Sample Requirements:

--- Customers can provide antigen and hybridoma cell lines directly.

--- Customers can provide antigens, hybridoma cell culture supernatants and TriZol lysates of hybridoma cell lines (usually 5 x 106 cells / 2 ml Trizol).

---Shipping condition: communicate with our technical support in advance