Earticle

초록

영어

Antibody fragments such as F(ab)’2 are advantageous over a whole antibody molecule in antigen-antibody binding studies because of its reduced non-specific binding, lower immunogenicity, and eliminated Fcassociated effector functions. Pepsin and/or papain are generally used to obtain Fab and F(ab)’2 [1]. But these enzymes have such disadvanages as
long reaction time, acidic pH condition, and complicated purification. FabRICATOR, an IdeS, is a cysteine proteinase that can specifically cleave the hinge region of IgG at a neutral pH to generate well-defined Fc and F(ab)’2 fragments within 30 min [2]. It was purchased from Genovis, Inc., Sweden. In this study, we evaluated the FabRICATOR’s cleavage performance and devised the successive recovery process to obtain Fc and F(ab)’2. Rabbit IgG (gamma globulin) was used as a model IgG. For 50 μg of rabbit IgG, 100 unit of FabRICATOR was used at 37℃, pH 6.6 for 30 min. The isolation process consisted of three steps: (1) an immobilized
metal affinity column to capture FabRICATOR by affinity interaction between the poly-histidine tag of the enzyme and the nickel domain, (2) Protein A column to recover Fc, and (3) SEC to obtain purified F(ab)’2. SDS-PAGE and SEC-HPLC (Biosep SEC-2000) results indicated the cleavage and separation were effective and successful. We plan to use F(ab)’2 for immunodiagnostics application replacing whole antibody and Fc for chemical fusion with other therapeutic protein for improved stability.

키워드

저자

• Yun Kyoung NOH [ Dept. of Bionanotechnology, Hanyang University, Ansan. ]
• Eun Kyu LEE [ Dept. of Bionanotechnology, Hanyang University, Ansan. ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576