| Background Antibodies play important roles in biological research and preparing medical products. Particularly, mouse monoclonal antibody has been a dominant tool in research, diagnostic and therapeutic areas for a long time. However, the new technology of generating rabbit monoclonal antibodies has offered an improved alternative of mouse monoclonal antibody. CD163, also known as M130, is a type I transmembrane protein belonging to group B of the cysteinerich scavenger receptor family. CD 163 is specifically expressed on monocytes and macrophages and is inducible by immunosuppressant glucocorticoids and IL10. CD163 mediates binding of bacteria by monocytes, leading to the release of inflammatory cytokines. It is essential for the circulatory clearance of hemoglobin-haptoglobin (Hb-Hp) complexes as well as free hemoglobin. It can also mediate monocyte-erythroblast adhesion and promote erythroblast expansion. CD 163 binds and internalizes the cytokine TWEAK, and the ratio of soluble CD163 to TWEAK in the plasma is elevated during atherosclerosis. Moreover, CD 163 has been reported to have important applications in a number of other diseases including autoimmune diseases, tumors, liver failure, etc. Objective To develop a specific and high affinity CD163 RabMab with multiple applications. Methods An unique immuogen for mouse CD163 was designed and CD 163 RabMab was generated by improved rabbit hybridoma technology. Anti-sera and hybridoma supernatants were screened by ELISA, WB, IHC-P and ICC.The best positive hybridoma was cloned and expressed as recombinant RabMAb for further characterization. Additional tests on reported negative cell lines or tissues were used to confirm its specificity and affinity. Results A total of 68 ELISA positive clones were generate and 8 of them can work on WB. A clear 150kDa target band can be detected in all the positive lysates. Furthermore, no signal was detected on CD 163-negative cell lines or tissue lysates reported by literatures.#23 clone is the best one which can be used in other species like human and rat, and it shows good tissue specificity in IHC-P and IHC-Fr. Therefore, this clone was used for further RabMab production. Conclusions We successfully developed a specific anti-CD 163 RabMab, which can be used for ELISA, WB, IHC-P and IHC-Fr. Besides the original species, it can also be used in human and rat. As we know, this antibody should be the anti-CD 163 RabMab with best quanlity in the current market and should have many important applications. |
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