ProMab Biotechnologies has nearly 2 decades of experience generating highly specific and potent monoclonal antibodies.
We have generated over 5000 hybridoma cell lines for our catalog and custom projects for clients from academic institutions to biopharma.
Starting from a hybridoma cell line, we offer the following services:
- Validation of the hybridoma cell line in terms of antibody production, antibody isotyping, and antigen-binding specificity;
- RNA extraction and reverse transcription;
- scFv sequencing by PCR amplification and subcloning of the variable domains;
- Full antibody sequencing by subcloning both variable and constant domains.
We provide a high-quality hybridoma sequencing service, also called hybridoma development. Hybridoma sequencing specifically refers to the sequencing of the variable regions of monoclonal antibodies being produced from hybridoma cell lines and will be required for the production of humanized antibodies, or for the authentication of a hybridoma cell line.
Total RNA is extracted from 100,000 hybridoma cells and using PCR methods, and the known non- variable flanking constant region sequences of respective antibody isotypes, the antibody variable regions (both heavy chain, VH, and light chain, VL) will be amplified for cloning and sequencing.
Degenerate primers are used to amplify the variable
regions of murine or rabbit heavy (VH) and
light (VL) chain antibody transcripts. Not
all hybridomas generate single VH and
VL chain transcripts. Multiple cloned
and sequenced VH and
VL chains are expressed in full-length
antibody plasmid
backbones, and resulting
VH-VL pairs are examined
for binding to the target antigen. Single-chain
variable fragments (scFv) are synthesized,
incorporating a bacterial pelB secretion signal,a
linker between the VH and
VL domains, and a C-terminal 6xHis
purification tag.
Figure 2. scFvs are expressed in E. coli, and binding of the purified scFvs to the target antigen is verified by ELISA
Antigen-antibody binding will subsequently be confirmed by transient expression of either single-chain antibodies (scFv) or whole IgGs in a mammalian expression system. Further testing will be performed to confirm that the recombinant scFv or whole IgG retains antigen binding affinity and specificity to its target molecule.
We have >1600 hybridoma clones are available to generate CAR-T cells.
Other Custom Services:
HYBRIDOMAS THAT PRODUCE ANTIBODY AGAINST CELL-SURFACE ANTIGEN | ||
---|---|---|
Catlog # | Description | Entrez Gene ID |
30630 | Mouse Monoclonal Antibody to EPCAM | 4072 |
30623 | Mouse Monoclonal Antibody to ROR2 | 4920 |
30622 | Mouse Monoclonal Antibody to MSTN | 2660 |
30594 | Mouse Monoclonal Antibody to CSF1R | 1436 |
30590 | Mouse Monoclonal Antibody to CD14 | 929 |
30585 | Mouse Monoclonal Antibody to CD14 | 929 |
30578 | Mouse Monoclonal Antibody to CD10 | 4311 |
30576 | Mouse Monoclonal Antibody to CD10 | 4311 |
30570 | Mouse Monoclonal Antibody to CD93 | 22918 |
30568 | Mouse Monoclonal Antibody to CD93 | 22198 |
30567 | Mouse Monoclonal Antibody to ITGAM | 3684 |
30565 | Mouse Monoclonal Antibody to IGFBP7 | 3490 |
30564 | Mouse Monoclonal Antibody to IGFBP7 | 3490 |
30560 | Mouse Monoclonal Antibody to MCAM | 4162 |
30552 | Mouse Monoclonal Antibody to EGFR | 1956 |
30551 | Mouse Monoclonal Antibody to EGFR | 1956 |
30550 | Mouse Monoclonal Antibody to CD22 | 933 |
30549 | Mouse Monoclonal Antibody to CD22 | 933 |
30528 | Mouse Monoclonal Antibody to ALCAM | 214 |
30527 | Mouse Monoclonal Antibody to IL3RA | 3563 |
30522 | Mouse Monoclonal Antibody to IL2RA | 3559 |
30519 | Mouse Monoclonal Antibody to CD24 | 100133941 |
30516 | Mouse Monoclonal Antibody to IL3RA | 3563 |
30512 | Mouse Monoclonal Antibody to ALCAM | 241 |
30507 | Mouse Monoclonal Antibody to FLT3 | 2322 |
30505 | Mouse Monoclonal Antibody to MCAM | 4162 |
30498 | Mouse Monoclonal Antibody to CSPG4 | 1464 |
30495 | Mouse Monoclonal Antibody to FGF4 | 2249 |
30494 | Mouse Monoclonal Antibody to FGF4 | 2249 |
30490 | Mouse Monoclonal Antibody to NRCAM | 4897 |
30486P | Mouse Monoclonal Antibody to EGFR mutant | 1956 |
30486A | Mouse Monoclonal Antibody to EGFR mutant | 1956 |
30485P | Mouse Monoclonal Antibody to ITGA2B | 3674 |
30485A | Mouse Monoclonal Antibody to ITGA2B | 3674 |
30480P | Mouse Monoclonal Antibody to CD33 | 945 |
30480A | Mouse Monoclonal Antibody to CD33 | 945 |
30476A | Mouse Monoclonal Antibody to ITGB4 | 3691 |
30449P | Mouse Monoclonal Antibody to TGFBR3 | 7049 |
Other Components of the CAR Platform: