Enhancement Effect of Porcine Inhibin Polyclonal Antibody on Superovulation in C57BL/6J Mice

doi:10.12300/j.issn.1674-5817.2025.088

Abstract

Abstract:

Objective To prepare rabbit anti-porcine inhibin polypeptide-keyhole limpet hemocyanin(KLH) conjugated polyclonal antibody and evaluate its effect on superovulation in C57BL/6J mice. Methods New Zealand white rabbits were immunized with a synthesized porcine inhibin polypeptide conjugated with KLH to produce anti-inhibin serum (AIS, i.e., inhibin polyclonal antibody). Female C57BL/6J mice received intraperitoneal injections of purified AIS in combination with pregnant mare serum gonadotropin (PMSG), followed by human chorionic gonadotropin (hCG) after 48 hours to induce superovulation. Oocytes obtained from superovulation were collected and counted 15 hours post-hCG administration, and the number of 2-cell embryos was assessed 24 hours after in vitro fertilization. Results AIS prepared by immunizing New Zealand White rabbits with KLH-conjugated porcine inhibin polypeptide was subjected to titer determination by indirect ELISA, showing titers reaching 1∶ 512 000. SDS-polyacrylamide gel electrophoresis analysis of ammonium sulfate-purified AIS revealed distinct 50 kDa and 25 kDa bands corresponding to the theoretical molecular weights of IgG antibody heavy and light chains, confirming successful production of porcine inhibin polyclonal antibody. Compared with conventional superovulation methods, AIS diluted 10-fold combined with PMSG significantly increased the number of oocytes obtained from superovulation in mice (P<0.05) by approximately 1.5-fold. Conclusion Porcine inhibin polyclonal antibody, as an improved superovulation reagent, can improve superovulation efficiency in C57BL/6J mice, and shows promising prospects for future applications.

Key words: Porcine anti-inhibin serum, C57BL/6J female mice, Superovulation, Oocyte number, Number of 2-cell embryos

CLC Number:

Q95-33

ZHAO He,ZHANG Tao,LI Li,et al. Enhancement Effect of Porcine Inhibin Polyclonal Antibody on Superovulation in C57BL/6J Mice[J]. Laboratory Animal and Comparative Medicine, 2026, 46(2): 271-278. DOI: 10.12300/j.issn.1674-5817.2025.088.

Figures/Tables 5

References 29

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稀释浓度 Dilution ratio	免疫前血清 Pre-immune serum	免疫后血清 Post-immunization rabbit serum	PBST对照 PBST control
1∶1 000	0.086	2.106	0.070
1∶2 000	0.096	2.083	0.068
1∶4 000	0.086	2.033	0.069
1∶8 000	0.087	2.005	0.071
1∶16 000	0.076	1.139	0.069
1∶32 000	0.060	1.187	0.067
1∶64 000	0.075	1.067	0.067
1∶128 000	0.082	1.008	0.066
1∶256 000	0.073	0.761	0.065
1∶512 000	0.071	0.548	0.067
1∶1 024 000	0.073	0.573	0.069
Blank	0.075	0.239	0.067

稀释浓度 Dilution ratio	免疫前血清 Pre-immune serum	免疫后血清 Post-immunization rabbit serum	PBST对照 PBST control
1∶1 000	0.086	2.106	0.070
1∶2 000	0.096	2.083	0.068
1∶4 000	0.086	2.033	0.069
1∶8 000	0.087	2.005	0.071
1∶16 000	0.076	1.139	0.069
1∶32 000	0.060	1.187	0.067
1∶64 000	0.075	1.067	0.067
1∶128 000	0.082	1.008	0.066
1∶256 000	0.073	0.761	0.065
1∶512 000	0.071	0.548	0.067
1∶1 024 000	0.073	0.573	0.069
Blank	0.075	0.239	0.067

组别 Group	卵母细胞总数 Total number of oocytes	2-细胞期胚胎总数 Total number of 2-cell embryos
A2-1	22.67±3.51	14.00±3.61
A2-2	52.67±3.06^abc	47.67±2.52^abc
P2	36.00±5.29	31.00±5.57
H2	63.00±6.00^a	47.67±5.51^a

组别 Group	卵母细胞总数 Total number of oocytes	2-细胞期胚胎总数 Total number of 2-cell embryos
A2-1	22.67±3.51	14.00±3.61
A2-2	52.67±3.06^abc	47.67±2.52^abc
P2	36.00±5.29	31.00±5.57
H2	63.00±6.00^a	47.67±5.51^a