Bolton-Hunter Reagent의 Analog를 이용하여 Bolton-Hunter방법에 의해 단백질을 Radiolabeling시킬 때의 최적조건을 찾는 방법의 개발 = Use of an Analog of Bolton-Hunter Reagent to Find Optimal Conditions in Radioiodinating Proteins by Bolton-Hunter Method

다국어 초록 (Multilingual Abstract)

We report a method to find optimal conditions in radioiodinating proteins by using an analog of Bolton-Hunter reagent, 7-hydroxy-4 methylcoumarin 3-acetic acid, succinimidyl ester(CASE) before radioiodinating the proteins by Bolton-Hunter method. Hen egg lysozyme(HEL) and a monoclonal anti-lysozyme antibody, HyHel-10 were used as a model system. When different pHs and (CASE)/(lysozyme) molar ratios were tried to conjugate lysozyme with CASE, we found that 1 CASE molecule was conjugated to a lysozyme molecule at pH between 6.0 and 7.0 and at (CASE)/(lysozyme) molar ratio below 5.0. The resulting CASE-lysozyme complex has al-most the same binding affinity against HyHel-10 as intact lysozyme has. To radioiodinate more lysozyme uniformly, we used mixture of radioactive (RBH) and nonradioactive (NBH) Bolton-Hunter reagent in 1 : 29 molar ratio to radioiodinate lysozyme with (BH(RBH+NBH))/(lysozyme) molar ratio 4.0 at pH = 7.0. Radioiodinated lysozyme has a little lower binding affinity against HyHel-10 than intact lysozyme has, and its binding constant against HyHel-10 is approximately 109 M-'.