INH表位多肽疫苗抗体间接ELISA测定方法的建立及优化

doi:10.16431/j.cnki.1671-7236.2016.08.031

Abstract

Abstract:

In this study, an indirect ELISA method was established to detect inhibin hormone (INH) epitope peptide vaccine antibody, it would provide oretical reference for the determination of Fine-wool sheep after active immune body INH epitope peptide vaccine antibody. On the basis of the predecessors, using indirect ELISA method to determinate serum INH epitope peptide antibody levels of sheep, and through control different experimental conditions to look for the best experimental conditions. Through explorating the experimental conditions, finally, the testing experiment conditions were determined, which was blocked solution with skimmed milk powder, INH and GnIH synthetic peptides dilution degrees for 20 000 times, the optimum reaction time was 60 min, the best color action time was 15 min. In this experiment, a kind of method to detection antibody in the body after INH active immune sheep was built, it would provide a reference for future research.

Key words: inhibin hormone; Fine-wool sheep; active immunization; reproductive hormone

CLC Number:

S852.4

ZHANG Ling-ling, YUE Yao-jing, FENG Rui-lin, LI Hong-feng, GUO Ting-ting, YUAN Chao, NIU Chun-e, LIU Jian-bin, SUN Xiao-ping, HAN Ji-long, LIU Shan-bo, YANG Bo-hui. Establishment and Optimization of INH Epitope Peptide Vaccine for Detection of Antibody[J]. , 2016, 43(8): 2156-2163.

References

[1] 茆达干,杨利国,姜勋平,等.抑制素α(1~32)基因免疫对大鼠卵泡发育和生殖激素的影响[J].中国农业科学,2003,36(12):1154-1159.
[2] 茆达干,杨利国,张志杰,等.抑制素基因免疫的免疫反应性[J].中国兽医学报,2006,26(3):292-295.
[3] 姜勋平,杨利国,刘桂琼,等.抑制素基因免疫对小鼠生殖的影响[J].中国兽医学报,2002,22(4):368-369.
[4] Cummmins J M.Seprm size body massandre production inmammals[J].Seprmcell,1983,114(4):395-398.
[5] Findlay J K,Clarke I J,Robertson D M.Inhibin concentrations in ovarian and jugularvenous plasmaand the relationship of inhibin with follicle-stimulating hormone and luteinizing hormone during the ovineestrous cycle[J].Endocrinology,1990,126(1):528-535.
[6] Tsutsui K.Anewkey neuro hormone control ling reproduction,gonadotrop-ininhibit oryhormone (GnIH):Biosynthesis,mode of action and functional significance[J].Progressin Neurobiology,2009,88(1):76-88.
[7] Bartlewski P M,Beard A P,Cook S J,et al.Ovarian antral follicular dynamics and their relationships with endocrine evariables throughout the estrous cycle in breeds of sheep differing in prolificacy[J].J Reprod Fertil,2011,115(1):111-124.
[8] Clarke I J,Qi Y,Puspita S I,et al.Evidence that RF-amide related peptide sare inhibitors of reproduction in mammals[J].Frontiersin Neuroendocrinology, 2009,30(3):371-378.
[9] Condon T P,Leipheimer R E,Curry J J.Preliminary evidence for a CNS site of action for ovarian inhibin[J].Life Sciences, 2010,32(15):1691-1698.
[10] Miller W L,Huang E S.Secretion of ovine luteinzing hormone in vitro:Differential positive control by 17-β estraiol and a preparation of porcine ovarian inhibin[J].Endocr,1985,117(3):907-901.
[11] Wrathall J E.Inhibin immunoneut ralization by antibodis raised against synthetic peptide sequences of inhibin α-subunit effect son gonadotrophin concentrations and ovulation rate in sheep[J].Endocrinal,1990,124:167-176.
[12] Leizhihat L Y.Recent studies of gonadotropin-inhibitory hormone[J].Animal Husbandry & Veterinary Medicine,2008,40(9):60-100.
[13] 李万宏,王春强,周虚.抑制素对动物生殖系统的影响及其在动物生产中的应用[J].中国畜牧兽医,2011,38(6):204-207.
[14] Ding H Z,Yu B S.Relationship between gonadotropin-inhibitory hormone and hypothalamus-pituitary-gonadalaxis[J].Progressin ModernBiomedicine,2010,10(23):4598-4600.
[15] Hinuma S,Shintani Y,Fukusumi S,et al.New neuro peptides containing carboxy-terminal RF-amide and the irreceptor in mammals[J].Nat Cell Biol,2000,2(10):703-708.
[16] Ukena K,Iwakoshi E,Minakata H,et al.A novel rat hypothalamic RFamide-related peptide identified by immune affinity chromate graphy and mass spectrometry[J].FEBS Lett,2002,512(1-3):255-258.
[17] 焦彩兰,储明星,王金玉.绵羊高繁殖力内分泌学机制的研究进展[J].中国畜牧兽医,2006,33(1):30-32.
[18] Hennies M,Voglmayr J K,Dietrich E,et al.Hormonal resonse of femal goat stoactive immunization against are combinant human inhibin subunit,and establishment of an enzyme linked immune sorbernt assay for caprine follicle stimulating hormone[J].Reprod Dom Anim,2001,36(2):65-71.
[19] Naqvi S M,Gulyani R.The effect of gonadotrophin releasing hormone and follicle stimulating hormone in conjunction with pregnant mareserum gonadotrophinon the super ovulatory response in crossbred sheep in India[J].Trop Anim Health Prod,1998,30(6):369-376.
[20] 王水莲,薛立群,邢朝芳,等.卵泡抑制素与GFP融合基因疫苗免疫对大鼠卵巢和生殖激素的影响[J].中国兽医学报,2012,32(2):309-313.
[21] 王水莲,冯建忠,李文平,等.非抗性筛选抑制素基因疫苗免疫免疫对奶牛产后子宫复旧的影响[J].湖南大学农业学报,2014,40(4):395-398.

Establishment and Optimization of INH Epitope Peptide Vaccine for Detection of Antibody

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Zhiliang, YANG Xixi, TIE Yanan, FU Xiaoyu, WANG Xinrong. Effects of Superovulation on Morphology and Hormone Levels of Reproductive Organs in Rabbits [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2250-2258.
[2]	JIA Yinhai, HUANG Mingguang, XU Wenwen, LI Fangfang, WEN Xinwang, LI Ming, YANG Xiurong, WU Zhuyue, JIANG Hesheng. Relationship Between Salivary Reproductive Hormones and Crystals with Follicular Development in Buffalo During Estrus [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 592-598.
[3]	Lina·Shaken, Baheti·Boday, YANG Hua, XIE Jingang, DONG Qiaoyan, Wurelihazi·Hazihan. Effects of Inhibin α on Reproductive Hormone Content in Kazakh Sheep by Active and Passive Immunity [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 658-667.
[4]	FAN Yushan, WU Junhui, GAO Fenglei, GUO Conghui, XIE Shaoyi, MENG Li, LI Li, ZHANG Shouquan, WEI Hengxi. Effects of Timed Artificial Insemination on Reproductive Performance and Reproductive Hormone in Multiparous Sows [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3726-3733.
[5]	ZHAO Xiaoyu, SHAERSHANBIEKE Abudilida, MA Chen, WU Yingping, WANG Junhua, LI Haiying. Effects of Dietary Crude Protein Level on Laying Raws,Reproductive Hormone Concentrations and Related Gene mRNA Expression in Reproductive Axis of Yili Geese [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1729-1738.
[6]	WANG Tao, LI Yurong, HE Yunmei, XU Fuxun, SUN Xuefeng, A Lamusi, SUN Chunhe, ZHAO Shengguo, LIU Bin. Effects of Estrus Induction on Lambing and Serum Index of Cashmere Goats in Non-breeding Season [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1819-1827.
[7]	WANG Chunhui, ZHANG Liping, ZHANG Changji. Study on the Supplementary Effect of Straw Complete Pellet Feeds for Gansu Alpine Fine-wool Sheep in Late Pregnancy [J]. , 2018, 45(9): 2455-2462.
[8]	CHEN Ling, HUANG Dapeng, WEI Guosheng. Effects of Nutrient Level During Late Pregnancy on Reproductive Performance and Blood Biochemical Indexes of Primiparous Sows [J]. , 2018, 45(8): 2190-2196.
[9]	LIU Xiaojun, ZHANG Lichun, HAN Linkai, CHU Xitong, CUI Chengdu, LU Cheng, JIN Haiguo, SUN Fuliang. Cloning, Expression and Bioinformatics Analysis of KAP2 Gene in Sheep [J]. , 2018, 45(6): 1454-1462.
[10]	ZHAO Guodong, ZANG Changjiang, WANG Gen, XIE Jinglong, YANG Jiantao, TANG Xuemei, ZHANG Juanli, YANG Kailun. Effects of Supplement with Letrozole and N-methyl-D-aspartate on Plasma Reproductive Hormone Levels of Ewes on the Basis of Melatonin Supplementation [J]. , 2018, 45(12): 3455-3462.
[11]	YU Shun-yu, LIU Kai-dong, LUAN Zhao-jin, WANG Guo-yi, LIU Nan, HE Jian-ning. Expression Analysis of TGF-βRⅠ Gene in Ovary of AoHan Fine-wool Sheep at Different Tissues and Estrous Cycles [J]. , 2017, 44(7): 1941-1946.
[12]	MA Shuang, SONG Peng-yan, ZHAI Fu-zhan, ZHONG Xiu-hui. Effects of Maternal Exposure to Bisphenol A on Survival Rate, Reproductive Hormones and Relative Genes of Offspring Mice [J]. , 2017, 44(7): 2033-2041.
[13]	CHENG Yuan, ZHAO Xiao-yu, LI Yuan-yuan, ZHAO Quan-zhuang, ZHU Jian-ping, AI Shan-jiang, LI Hai-ying. Study on the Changes of Reproductive Hormone and Regulation of Follicle Development During the Reproductive Cycle of Yili Goose in Xinjiang [J]. , 2017, 44(11): 3264-3269.
[14]	FU Xue-feng, YANG Han-yulu, SHI Gang, XU Xin-ming, ZHANG En-ping, DI Jiang, ZHANG Yan-hua, HUANG Xi-xia, TIAN Ke-chuan. Proteome Array on Differentially Expressed Proteins of Skin Tissue in Fine-wool Sheep with Different Fiber Diameter [J]. , 2016, 43(4): 879-891.
[15]	ZHANG Ling-ling, YANG Bo-hui, YUE Yao-jing, GUO Ting-ting, LIU Jian-bin, YUAN Chao, NIU Chun-e, FENG Rui-lin, GUO Jian, SUN Xiao-ping, LIU Shan-bo. Effects of Active Immunization with GnIH and INH Epitope Peptide Vaccine on Reproductive Hormones of Gansu Alpine Fine-wool Sheep [J]. , 2016, 43(4): 1039-1044.