Isolation and Characterization of Antibody Induced by 71 kDa Bovine Protein (Anti-P71) from Pregnant Cow Serum for Detection of Pregnancy in Cattle
Mazin Amir Mohammed Khlil
, Aulanni`am Aulanni`am , Arie Srihardyastutie , Tiara Chintya Devi , Hanna Merryani Suwanto
indirect ELISA, Cows Pregnancy, PAG71, Western Blot, PSP-B,
Abstract—The efficiency of animal production is important from a physical and economic point of view for livestock breeders, and reproduction is considered an important matter. Therefore, detecting the occurrence of pregnancy early after mating in cattle is very important to ensure the success of pregnancy and the preservation of the fetus until the birth of the new offspring, and confirming the occurrence of pregnancy as soon as possible is very important to preserve the sperm. Remarriage after pregnancy leads to the death of the fetus. Therefore, the main objective of this study was to isolate Pregnancy Associated Glycoproteins and use it to detect pregnancy in cattle in the shortest possible time after mating. The bp71 kDa protein was isolated using "SDS-PAGE" where the proteins are separated depending on the difference in molecular weight, and then the Pregnancy Associated Glycoproteins was extracted from the gel using the electroelution technique. After extracting the Pregnancy Associated Glycoproteins mixed with adjuvants and then injected into experimental animals to stimulate the immune system of those rabbits, which in turn produce antibodies to the Pregnancy Associated Glycoproteins of cows, and fore confirm the formation of antibodies, the indirect ELISA technique was used to determine the amount Antibodies formed and the time period required for the highest production of antibodies. Also, Western blot technique was used to ensure the ability of the antibodies to identify the Pregnancy Associated Glycoproteins bp71 kDa. the antigen used was PAG71. The primary antibody used was PAG71 antibody induced by PAG71 derived from rabbit serum, while the secondary antibody used was anti-Rabbit AP Conjugated. the results of serum protein bands in pregnant cows, protein 71 kDa was found in the serum of pregnant cows aged 2 months, 5 months, 6 months, and 7 months, and in the serum of non-pregnant cows there was no protein 71.the result of western blot protein bands with a molecular weight of 71 kDa were seen in a sample of 7-month-old pregnant cows, while in non-pregnant cattle samples, no protein was found on protein bars of 71 kDa molecular weight. Protein bands with a molecular weight of 71 kDa were seen in a sample of 7-month-old pregnant cows, whereas in non-pregnant cattle samples, no protein bands of this molecular weight were found. In Conclusion, SDS-PAGE confirms the association of the protein PAG with cattle pregnancy, as mentioned in previous studies, and the Pregnancy Associated Glycoproteins appears in the range of 50 to 100 kDa. in western blot technique, correlation was observed Antibodies to pregnancy-associated protein in the serum of pregnant cows did not appear in the serum of non-pregnant cows.
"Isolation and Characterization of Antibody Induced by 71 kDa Bovine Protein (Anti-P71) from Pregnant Cow Serum for Detection of Pregnancy in Cattle", IJSDR - International Journal of Scientific Development and Research (www.IJSDR.org), ISSN:2455-2631, Vol.8, Issue 7, page no.728 - 733, July-2023, Available :https://ijsdr.org/papers/IJSDR2307105.pdf
Volume 8
Issue 7,
July-2023
Pages : 728 - 733
Paper Reg. ID: IJSDR_207788
Published Paper Id: IJSDR2307105
Downloads: 000347268
Research Area: Chemistry
Country: Kota malamg, Jawa Timur, Indonesia
DOI: http://doi.one/10.1729/Journal.35210
ISSN: 2455-2631 | IMPACT FACTOR: 9.15 Calculated By Google Scholar | ESTD YEAR: 2016
An International Scholarly Open Access Journal, Peer-Reviewed, Refereed Journal Impact Factor 9.15 Calculate by Google Scholar and Semantic Scholar | AI-Powered Research Tool, Multidisciplinary, Monthly, Multilanguage Journal Indexing in All Major Database & Metadata, Citation Generator
Publisher: IJSDR(IJ Publication) Janvi Wave
