Production of recombinant monoclonal antibodies against HSET

HSET (Human Spleen Embryo Testis), belonged to the kinesin 14A family, is a protein required for centrosome clustering during cell division. Since cancer cells with extra-centrosomes could lead to multi-polar division, resulting in genomic instability and cell death, these cancer cells depend on the centrosome clustering activity of HSET that helps normal chromosome segregation by focusing several centrosomes into two poles. As the first step of the strategy to inhibit the function of HSET in cancer cells, we produced mouse monoclonal antibodies (mAbs) against HSET using a hybridoma technique and characterized them. Peptide antigens of 12 amino acid (aa)-length, designed from the regions corresponding to N-terminus (1-134th aa) and C-terminus (626-672th aa) that do not have homology with other kinesin molecules, were used for mouse immunization. Total 28 of mAbs against the peptides from N-terminus HSET were obtained, but no mAb against C-terminus of HSET. In ELISA with the Abs in ascitic fluids against yeast surface displayed HSET, clone 8C346 showed high affinity. In ELISA using Abs purified from hybridoma culture supernatants, clone 1C274, 2C280, 2C281, 6C407, 9C352, and 9C353 showed relatively strong binding to Whole HSET protein. We expect that Abs with high binding to HSET could inhibit HSET function, resulting in selective death of cancer cells.

Author dc:creator

• 서, 영실

Contributors dc:contributor

• 권, 명희
• 대학원 의생명과학과
• 112605

• HSET
• Centrosome clustering
• Anti-HSET antibody
• Hybridoma
• Cancer
• 중심체 군집
• 항-HSET 항체
• 하이브리도마
• 암

Language dc:language

en

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000017694
000000017694

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/10863