Objective: To prepare and calibrate the first batch national standard(NS) of recombinant human chorionic gonadotrophin (rHCG) for bioassay. Methods: The bioactivity of the first batch of the candidate preparation of rHCG was collaboratively calibratedby five laboratories taking the fifth batch of rHCG of WHO international standard 07/364 with a titer of 162 IU per vial as the standard, the 1209 bioassay method of rHCG in Volume Ⅳof the Chinese Pharmacopoeia was adopted. The structure of candidate preparation concentrate was confirmed by imaging capillary isoelectric focusing(icIEF), peptide mapping and determination of relative molecular mass. The key physicochemical properties were measured, such as the protein content by ultraviolet spectrophotometry, purity and polymers by SEC-HPLC, oxidation of subunits by RP-HPLC. The stability tests for the candidate preparation were carried out. Results:The biopotency of the candidate preparation of rHCG was finally determined to be 6 400 IU per vial after collaborative calibration. The content of rHCG was about 290 μg per vial, and the purity by SEC-HPLC was about 100%. Structure confirmation results meet the requirements; Physical and chemical indicators such as polymer, dissociated subunit, and oxidized subunits were conformed to the standard; the stability test results meet the requirements. Conclusion: The candidate preparation is recognized as rHCG to sever as the 1st NS rHCG for bioassay of rHCG and related preparations, and its biopotency is 6 400 IU per vial.
LI Zhan-jun, LIANG Yu-ling, LI Yi, HU Xin-yue, HU Yu-chi, CAO Chun-ran, WEI Xia, ZHU Qing-fen, ZHANG Juan, FENG Run-dong, WANG Li-fang, HE Li-xiu, LI Jing, ZHANG Hui, LIANG Cheng-gang
. Preparation and collaborative calibration of the 1st national standard of recombinant human chorionic gonadotrophin for bioassay*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(1)
: 68
-77
.
DOI: 10.16155/j.0254-1793.2022.01.08
[1] LATHI RB, MILKI AA. Recombinant gonadotropins[J].Curr Womens Health Rep, 2001, 1(2):157
[2] LOUMAYE E, MARTINEAU I, PIAZZI A, et al. Clinical assessment of human gonadotrophins produced by recombinant DNA technology[J].Hum Reprod, 1996, 11(Suppl 1):95
[3] GRAY CJ. Glycoprotein gonadotropins. Structure and synthesis[J].Acta Endocrinol Suppl (Copenh), 1988, 288: 20
[4] BELLAVIA M, de GEYTER C, STREULI I, et al. Randomized controlled trial comparing highly purified(HP-hCG) and recombinant hCG(r-hCG) for triggering ovulation in ART[J].Gynecol Endocrinol, 2013, 29(2):93
[5] CHOI J, SMITZ J. Luteinizing hormone and human chorionic gonadotropin: origins of difference[J].Mol Cell Endocrinol, 2014, 383(1-2):203
[6] BTELFELD AP, POUR SJ, POSCHMANN G, et al. A proteome approach reveals differences between fertile women and patients with repeated implantation failure on endometrial level does hCG render the endometrium of RIF patients?[J].Int J Mol Sci, 2019, 20(2):425
[7] SIMOPOULOU M, SFAKIANOUDIS K, MAZIOTIS E, et al. Investigating the optimal time for intrauterine human chorionic gonadotropin infusion in order to improve IVF outcome: a systematic review and meta-analysis[J].In Vivo, 2019, 33(6):1737
[8] DRISCOLL GL, TYLER JP, HANGAN JT, et al. A prospective, randomized, controlled, double-blind, double-dummy comparison of recombinant and urinary HCG for inducing oocyte maturation and follicular luteinization in ovarian stimulation[J].Hum Reprod, 2000, 15(6):1305
[9] KIM YJ. Glucocorticoid therapy in assisted reproduction[J].Clin Exp Reprod Med, 2021, 48(4):295
[10] GAO M, JIANG X, LI B, et al. Intrauterine injection of human chorionic gonadotropin before embryo transfer can improve in vitro fertilization-embryo transfer outcomes: a meta-analysis of randomized controlled trials[J].Fertil Steril, 2019, 112(1):89
[11] MAKRIGIANNAKIS A, VREKOUSSIS T, ZOUMAKIS E, et al. the role of HCG in implantation: a mini-review of molecular and clinical evidence[J].Int J Mol Sci, 2017, 18(6):1305
[12] ANTOINE JM, LOUMAYE E, ALVAREZ S, et al. Clinical use of recombinant gonadotropins(FSH, LH, hCG)[J].Contracept Fertil Sex, 1997, 25(2):141
[13] FONJALLAZ P, LOUMAYE E. Recombinant hCG(OVIDREL) and recombinant interferon-(beta)1a (REBIF). (No. 13 in a series of articles to promote a better understanding of the use of genetic engineering)[J].J Biotechnol, 2001, 87(3):279
[14] CHANG P, KENLEY S, BURNS T, et al. Recombinant human chorionic gonadotropin(rhCG) in assisted reproductive technology: results of a clinical trial comparing two doses of rhCG(Ovidrel) to urinary hCG(Profasi) for induction of final follicular maturation in in vitro fertilization-embryo transfer[J].Fertil Steril, 2001, 76(1):67
[15] 毕星宇, 刘俊芬, 武学清. 胚胎移植前宫腔内注射艾泽在IVF-ET中的应用[J].中国医药指南, 2016, 14(19):1
BI XY, LIU JF, WU XQ. The application of intrauterine injection before embryo transfer in IVF-ET eiser[J].Guide China Med, 2016, 14(19):1
[16] 何荣华, 高丽婷, 朱红雨, 等. 重组人绒毛膜促性腺激素研究进展[J].药物生物技术, 2018, 25(6):542
HE RH, GAO LT, ZHU HY, et al. Research advances of recombinant human chorionic gonadotropin[J].Pharm Biotechnol, 2018, 25( 6):542
[17] 李湛军, 李晶, 张慧, 等. 第1批重组人促卵泡素生物测定用国家标准品的制备与标定[J].药物分析杂志, 2018, 27(12):1350
LI ZJ, LI J, ZHANG H, et al. Preparation and calibration of the 1st batch national standard of recombinant human follicle-stimulating hormone for bioassay[J].Chin J Pharm Anal, 2018, 27(12):1350
[18] 王军志. 生物技术药物研究开发和质量控制[M].第3版. 北京: 科学出版社, 2018: 730
WANG JZ. Research, Development and Quality Control of Biopharmaceuticals[M].3rd. Beijing: Science Press, 2018: 730
[19] 周海钧. 生物统计方法[M].北京: 人民卫生出版社, 1983
ZHOU HJ. Biostatitical Methods[M].Beijing: People’s Medical Publishing House, 1983
