Anti-ceacam5 antibody and use thereof
An antibody, CDR1-H technology, applied in the direction of antibodies, anti-tumor drugs, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve the problem of poor internalization of surface proteins
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0536] Example 1: Preparation of recombinant extracellular domain of CEACAM protein
[0537] In this example, extracellular CEACAM derived from human (h) or cynomolgus (c) has been prepared by transient expression in human embryonic kidney HEK293 cells using plasmids expressing each cDNA as outlined in Table 1 protein domain (ECD).
[0538] Each expression plasmid is combined with 293fectin TM (Life Technologies) complexed and added to suspension cultured 293-F cells (derived from HEK293 cells). On day 8 post-transfection, culture supernatants were collected and the corresponding soluble proteins were purified by IMAC (GE Healthcare) to generate protein batches (see Table 1).
[0539] Table 1 :Description of the recombinant extracellular domain of the CEACAM protein
[0540]
Embodiment 2
[0541] Example 2: Generation of mouse anti-CEACAM5 monoclonal antibody
[0542] In this example, monoclonal antibodies were produced following immunization of mice according to a protocol that produces anti-CEACAM5 mAbs.
Embodiment 21
[0543] Example 2.1: Immunization and Hybridoma Generation
[0544] Immunization, fusion and screening using P3X63-Ag8.653 myeloma cells with the extracellular domain of human CEACAM5, the extracellular domain of cynomolgus monkey CEACAM5 or using e.g. Wennerberg A.E et al, 1993. Am. J. Pathol., 143(4), 1050-1054 and human UMC11 tumor cells as described in Kilpatrick et al, 1997. Hybridoma 16:381389.
[0545] 6-8 week old female BALB / c mice (S082342; Charles River Labs, Bar Harbor, ME) were bred at 3-4 day intervals using the RIMMS method as described by Kilpatrick et al (1997. Hybridoma 16:381389). Each received four rounds of immunizations over the course of 14 days. Antigen emulsified in Titermax adjuvant (TierMax Gold adjuvant; Sigma No. T2684) was administered subcutaneously to six sites along the back of the mice adjacent to draining lymph nodes and six side-by-side sites along the abdomen. Mice were sacrificed on day 4 after the last injection. Bilateral popliteal, ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More