WO2023080500A1 - Anticorps de diagnostic de grossesse bovine et kit de diagnostic de grossesse précoce l'utilisant - Google Patents

Anticorps de diagnostic de grossesse bovine et kit de diagnostic de grossesse précoce l'utilisant Download PDF

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Publication number
WO2023080500A1
WO2023080500A1 PCT/KR2022/016087 KR2022016087W WO2023080500A1 WO 2023080500 A1 WO2023080500 A1 WO 2023080500A1 KR 2022016087 W KR2022016087 W KR 2022016087W WO 2023080500 A1 WO2023080500 A1 WO 2023080500A1
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val
arg
pregnancy
amino acid
acid sequence
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PCT/KR2022/016087
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English (en)
Korean (ko)
Inventor
이재호
이상진
조용범
김수경
구자령
이재준
신상우
백다희
Original Assignee
주식회사 나노바이오라이프
이재호
이상진
조용범
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Application filed by 주식회사 나노바이오라이프, 이재호, 이상진, 조용범 filed Critical 주식회사 나노바이오라이프
Publication of WO2023080500A1 publication Critical patent/WO2023080500A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/689—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to pregnancy or the gonads

Definitions

  • the present invention relates to a bovine pregnancy diagnosis antibody and its use, and relates to the development of a bovine pregnancy diagnosis antibody using a specific part of bovine bPAG protein as an antigen and a bovine early pregnancy diagnosis kit using the developed antibody.
  • Cows have an estrous cycle of 21 days, and during estrus, sperm and eggs meet in the fallopian tube by artificial insemination or natural mating, go through cleavage, and then fertilize eggs between 9 and 13 days after fertilization. It implants in the wall of the uterus and becomes pregnant. A cow usually gives birth after a gestation period of 280-285 days.
  • non-pregnant cows can be re-fertilized during the next estrus, that is, after 21 days of fertilization. Then, the economic loss of farmers can be minimized. However, if pregnancy is not accurately determined at this time, re-fertilization should be done about 42 days after the start of the second estrus cycle or about 63 days after the start of the 3rd estrus cycle. If non-pregnancy continues for such a long period of time, the economic loss to farmers is enormous.
  • cow pregnancy diagnoses are performed by veterinarians or artificial insemination technicians between 35 and 60 days of pregnancy by fetal membrane stimulation (rectal examination) or ultrasound pregnancy diagnosis, and another method is radioimmunoassay using progesterone in milk or blood. , RIA).
  • Progesterone is a steroid hormone secreted by the corpus luteum and necessary for the normal progress and maintenance of pregnancy. It is a non-pregnancy diagnosis method using the principle that the concentration of progesterone in blood or milk increases when a cow is pregnant, and the concentration of progesterone decreases in the case of a non-pregnant cow.
  • these diagnostic methods have several problems.
  • the first is the use of radioactive isotopes.
  • EPF early pregnancy factor
  • ECG equine chorionic gonadotrophin
  • bPL placental lactogen
  • placental glycoprotein placental glycoprotein
  • placental glycoprotein placental glycoprotein
  • bovine pregnancy-associated glycoprotein bovine pregnancy.
  • An object of the present invention is to provide a diagnostic kit with improved diagnostic ability of bovine pregnancy by using a specific part of bovine bPAG protein as an antigen.
  • the present invention provides a protein for diagnosing bovine pregnancy that is expressed in the blood of pregnant cows.
  • the present invention provides a protein for diagnosing bovine pregnancy characterized in that the amino acid sequence 1 below.
  • the present invention provides a protein for diagnosing bovine pregnancy, characterized in that it has the amino acid sequence 2 below.
  • the present invention provides an antibody for diagnosing bovine pregnancy using amino acid sequence 1.
  • the present invention provides an antibody for diagnosing bovine pregnancy using amino acid sequence 2.
  • the present invention provides a kit for diagnosing bovine pregnancy using the antibody for diagnosing bovine pregnancy.
  • Figure 1 shows a comparison between the accuracy of the diagnostic kit according to the present invention and the existing diagnostic kit for bovine pregnancy.
  • a protein for diagnosing bovine pregnancy expressed in the blood of a pregnant cow is provided.
  • a protein for diagnosing bovine pregnancy characterized in that the amino acid sequence 1 below is provided.
  • the present invention provides a protein for diagnosing bovine pregnancy, characterized in that it has the amino acid sequence 2 below.
  • an antibody for diagnosing bovine pregnancy using amino acid sequence 1 is provided.
  • an antibody for diagnosing bovine pregnancy using amino acid sequence 2 is provided.
  • kits for diagnosing bovine pregnancy using the antibody for diagnosing bovine pregnancy is provided.
  • bovine pregnancy-associated glycoprotein (bPAG) is as follows.
  • the primary antigen sequence selected from the bPAG amino acid sequence is as follows, and Cys was added to the N-terminus for conjugation.
  • the secondary antigen sequence selected from the bPAG amino acid sequence is as follows, and Cys was added to the N-terminus for conjugation.
  • Fig. 1 shows the results of a test for diagnosis of gonorrhea in cattle using the primary and secondary antigen sequences.
  • the sample (A) judged positive by the existing diagnostic kit (K) appeared negative in the test (B) using the antigen according to the present invention, and as a result of the determination using ultrasound, the cow was confirmed as non-pregnant As a result, it was confirmed that the present invention has a higher accuracy for determining a cow's pregnancy than conventional diagnostic kits.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Pregnancy & Childbirth (AREA)
  • Reproductive Health (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne un anticorps de diagnostic de grossesse bovine, qui utilise une partie spécifique d'une protéine bPAG bovine en tant qu'antigène, et son utilisation. En particulier, en fournissant un kit de diagnostic de grossesse bovine ayant une plus grande précision que les produits classiques, la présente invention peut réduire l'intervalle de vêlage par raccourcissement de la période d'inter-grossesse, ce qui permet de réduire les coûts d'alimentation, les coûts d'exploitation, les coûts de main-d'œuvre, et les frais généraux de l'élevage de bovins, ainsi que d'améliorer l'efficacité de la production de veaux de 3 têtes par 5 ans classiquement à 4 têtes par 5 ans, et peut ainsi augmenter le revenu des exploitations bovines.
PCT/KR2022/016087 2021-11-04 2022-10-20 Anticorps de diagnostic de grossesse bovine et kit de diagnostic de grossesse précoce l'utilisant WO2023080500A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020210150843A KR20230065047A (ko) 2021-11-04 2021-11-04 소 임신 진단 항체와 이를 이용한 조기 임신 진단키트 개발
KR10-2021-0150843 2021-11-04

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WO2023080500A1 true WO2023080500A1 (fr) 2023-05-11

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003043524A2 (fr) * 2001-11-20 2003-05-30 The Curators Of The University Of Missouri Compositions et methodes pour le diagnostic precoce precis de la grossesse
US6869770B1 (en) * 1998-03-20 2005-03-22 The Curators Of The University Of Missouri Compositions and methods for early pregnancy diagnosis
KR20100110818A (ko) * 2007-12-13 2010-10-13 몬산토 테크놀로지 엘엘씨 조기 임신 진단을 위한 조성물 및 방법
KR20190055741A (ko) * 2017-11-15 2019-05-23 (주)에스엘에스바이오 bPAG1에 특이적으로 결합하는 항체 및 이의 용도

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6869770B1 (en) * 1998-03-20 2005-03-22 The Curators Of The University Of Missouri Compositions and methods for early pregnancy diagnosis
WO2003043524A2 (fr) * 2001-11-20 2003-05-30 The Curators Of The University Of Missouri Compositions et methodes pour le diagnostic precoce precis de la grossesse
KR20100110818A (ko) * 2007-12-13 2010-10-13 몬산토 테크놀로지 엘엘씨 조기 임신 진단을 위한 조성물 및 방법
KR20190055741A (ko) * 2017-11-15 2019-05-23 (주)에스엘에스바이오 bPAG1에 특이적으로 결합하는 항체 및 이의 용도

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE Protein NCBI; 19 March 2015 (2015-03-19), ANONYMOUS : "Pregnancy-associated glycoprotein 1, partial [Bos mutus]", XP093064522, Database accession no. ELR57007.1 *
ZOLI AP, GUILBAULT LA, DELAHAUT P, ORTIZ WB, BECKERS JF: "Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis", BIOLOGY OF REPRODUCTION, NEW YORK, NY [U.A.] : ACADEM. PRESS, US, vol. 46, no. 01, 1 January 1992 (1992-01-01), US , pages 83 - 92, XP002109207, ISSN: 0006-3363, DOI: 10.1095/biolreprod46.1.83 *

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