CAJ | 학술논문

目的：建立人促卵泡激素（FSH）体外生物学活性检测方法，并对方法进行验证。方法：以人卵巢颗粒细胞KGN为基质细胞，用不同浓度的FSH与KGN细胞表面的FSH受体结合，刺激细胞产生不同浓度的孕酮，通过测定细胞培养上清中的孕酮含量来评价FSH的生物学活性，结果用国际标准品进行校正；对方法的专属性、线性与范围、回收率、精密度、耐用性进行验证；对市售FSH产品进行检测，并与传统动物体内活性检测结果进行比较。结果：KGN细胞对FSH有很好的剂量反应关系，方法专属性符合要求；线性范围为0.1~200 ng/mL，相关系数R2≥0.99；回收率为80%~120%；经3次重复测定，3批市售样品和3批自制样品的活性分别为（13.4±0.4）~（13.5±0.8）和（12.9±0.7）~（14.3±1.2） IU/μg，变异系数在15%之内；结果显示该方法有很好的耐用性。测定3批市售重组人FSH产品和1批尿源FSH国家标准品，其体外活性测定结果与传统动物体内活性测定结果一致。结论：建立并验证了一种利用KGN细胞体外测定FSH生物学活性的方法，有望代替传统的动物体内活性测定方法，可用于FSH的生物学活性评价和质量控制。
목적：건립인촉란포격소（FSH）체외생물학활성검측방법，병대방법진행험증。방법：이인란소과립세포KGN위기질세포，용불동농도적FSH여KGN세포표면적FSH수체결합，자격세포산생불동농도적잉동，통과측정세포배양상청중적잉동함량래평개FSH적생물학활성，결과용국제표준품진행교정；대방법적전속성、선성여범위、회수솔、정밀도、내용성진행험증；대시수FSH산품진행검측，병여전통동물체내활성검측결과진행비교。결과：KGN세포대FSH유흔호적제량반응관계，방법전속성부합요구；선성범위위0.1~200 ng/mL，상관계수R2≥0.99；회수솔위80%~120%；경3차중복측정，3비시수양품화3비자제양품적활성분별위（13.4±0.4）~（13.5±0.8）화（12.9±0.7）~（14.3±1.2） IU/μg，변이계수재15%지내；결과현시해방법유흔호적내용성。측정3비시수중조인FSH산품화1비뇨원FSH국가표준품，기체외활성측정결과여전통동물체내활성측정결과일치。결론：건립병험증료일충이용KGN세포체외측정FSH생물학활성적방법，유망대체전통적동물체내활성측정방법，가용우FSH적생물학활성평개화질량공제。
Objective: To develop and verify a novel in vitro assay for determination of biological activity of hu?man follicle stimulating hormone(FSH). Methods: The assay is based on a human ovary granulosa cell line, KGN cell line, which expresses the human FSH receptor on its surface. FSH binding can stimulate KGN cell to express and secret progesterone, concentration of which is determined by a commercially available ELISA kit. The devel?oped method was verified for specificity, linear range, recovery, precision, and tolerance. Three batches of commer?cially recombinant FSH products and one batch of Urofollitropin were assayed by the method, and the results were compared with the results of traditional in vivo assay. Results: Highly dose-dependent progesterone secretion was observed at 72 h of incubation for KGN cell. The method met the specificity requirement. The linear range was 0.1~200 ng/mL, R2≥0.99. The recovery was within 80%~120%, which met the recovery requirement in activity assay of biological products. The results of three batches of commercial FSH products and three bathes of samples were (13.4±0.4)~(13.5±0.8) and (12.9±0.7)~(14.3±1.2) IU/μg respectively, CV%≤15%. The results showed good toler?ance to different progesterone ELASA kits. Furthermore, it was demonstrated that the assay delivered comparable re?sults to the in vivo assay for three batches of commercially recombinant FSH products and one batch of Urofollitrop?in. Conclusion: The new in vitro assay for FSH activity is developed and verified, which is supposed to replace the in vivo assay in the future and can be used for the biological activity assessment and quality control for FSH.