Earticle

초록

영어

Previously we reported novel target-specifuc binding proteins of modest potency that was derived from a synthetic kringle domain(KD) library on the yeast cell surface. We isolated KD variants that bind to anti-cancer target proteins, such as human death receptor 4(DR4) and/or DR5 and human tumor necrosis factor-a (TNFa). We selected five KD variants (KD413, KD415, KD506, KD548 and KDT26) which have specific biological activity. The anti-DR4 KD variants (KD413 and KD415) and anti-DR5 KD variants (KD506 and KD548) function as agonists to induce
apoptotic cell death in several cancer cell lines in vitro and antagonistic KD variant that bind to TNFa (KDT26) neutralizes TNFa-induced cytotoxicity. However, the selected KD variants have low affinity (KD =~10-7M) compare with TRAIL and infliximab. For affinity improvement,
we engineered loops of KD variants particularly loop5 and loop6 which are selected by loop mapping by yeast surface display. Using yeast surface display system, libraries were constructed to minimize the number of non-viable structures by rational design of nucleotide mixtures which bias for the wild-type (parent mutant) residues. The high affinity mutants
of the KD variants were isolated by using MACS and FACS systems. Kinetic analysis of KD variants from isolated mutants revealed that the affinity was improved. In the conference, we will present the relationship between the affinity and biological activity.

키워드

저자

• Chang Goo KIM [ Department of Molecular Science and Technology, Ajou University ]
• Chang Han LEE [ Department of Molecular Science and Technology, Ajou University ]
• Yong Sung KIM [ Department of Molecular Science and Technology, Ajou University ]

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간행물

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• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576