Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
Molecular Medicine Reports
Join Editorial Board Propose a Special Issue
Print ISSN: 1791-2997 Online ISSN: 1791-3004
Journal Cover
June-2017 Volume 15 Issue 6

Full Size Image

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
June-2017 Volume 15 Issue 6

Full Size Image

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article Open Access

Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography

  • Authors:
    • Shuangshuang Liu
    • Shasha Li
    • Yang Zhang
    • Ye Wang
    • Yumeng Zhu
    • Bin Wang
    • Zhi‑Nan Chen
  • View Affiliations / Copyright

    Affiliations: Department of Cell Biology, National Translational Science Center for Molecular Medicine, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 4035-4040
    |
    Published online on: April 27, 2017
       https://doi.org/10.3892/mmr.2017.6523
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

The immunoglobulin superfamily member CD147 is a widely expressed glycoprotein that occurs in both a membrane‑spanning and soluble form. Sandwich ELISA is a powerful tool for analyzing soluble antigens. The aim of the present study was to obtain a highly specific polyclonal antibody against human CD147 that can be used for sandwich ELISA analysis. Expression of recombinant CD147 by a eukaryotic expression system was used to immunize rabbits to obtain antiserum. A highly specific polyclonal antibody that was able to detect soluble CD147 in sandwich ELISA was obtained by antigen‑immunoaffinity chromatography purification. The purity of rabbit anti‑CD147 polyclonal antibodies was ~99%, and ELISA analysis was able to determine the titer of the rabbit anti‑CD147 polyclonal antibodies at 1:512,000. The lowest concentration of the standard CD147 antigen that the sandwich ELISA was able to detect was 31.25 pg/ml. The sandwich ELISA system was composed of anti‑hepatoma HAb18 monoclonal antibodies and purified rabbit anti‑CD147 polyclonal antibodies. The present study demonstrated that antigen‑immunoaffinity chromatography may be a good technique for the purification of polyclonal antibodies, which may be used to detect antigen in sandwich ELISAs.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

View References

1 

Biswas C: Tumor cell stimulation of collagenase production by fibroblasts. Biochem Biophys Res Commun. 109:1026–1034. 1982. View Article : Google Scholar

2 

Ellis SM, Nabeshima K and Biswas C: Monoclonal antibody preparation and purification of a tumor cell collagenase-stimulatory factor. Cancer Res. 49:3385–3391. 1989.

3 

Polette M, Gilles C, Marchand V, Lorenzato M, Toole B, Tournier JM, Zucker S and Birembaut P: Tumor collagenase stimulatory factor (TCSF) expression and localization in human lung and breast cancers. J Histochem Cytochem. 45:703–709. 1997. View Article : Google Scholar

4 

Tang J, Wu YM, Zhao P, Yang XM, Jiang JL and Chen ZN: Overexpression of HAb18G/CD147 promotes invasion and metastasis via alpha3beta1 integrin mediated FAK-paxillin and FAK-PI3K-Ca2+ pathways. Cell Mol Life Sci. 65:2933–2942. 2008. View Article : Google Scholar

5 

Wu Y, Zhou X and Zheng PS: Involvement of CD147 isoform-4 in the proliferation of SiHa cells: A possible molecular mechanism of cervical cancer. Oncol Rep. 26:717–724. 2011.

6 

Chen ZN, Yang Z and Mi L: Analysis on the structure and function of hepatoma transfer-associated factor HAb18G (in Chinese). J Mol Cell Immunol. 15:341999.

7 

Jiang JL, Zhou Q, Yu MK, Ho LS, Chen ZN and Chan HC: The involvement of HAb18G/CD147 in regulation of store-operated calcium entry and metastasis of human hepatoma cells. J Biol Chem. 276:46870–46877. 2001. View Article : Google Scholar

8 

Haug C, Lenz C, Diaz F and Bachem MG: Oxidized low-density lipoproteins stimulate extracellular matrix metalloproteinase Inducer (EMMPRIN) release by coronary smooth muscle cells. Arterioscler Thromb Vasc Biol. 24:1823–1829. 2004. View Article : Google Scholar

9 

Egawa N, Koshikawa N, Tomari T, Nabeshima K, Isobe T and Seiki M: Membrane type 1 matrix metalloproteinase (MT1-MMP/MMP-14) cleaves and releases a 22-kDa extracellular matrix metalloproteinase inducer (EMMPRIN) fragment from tumor cells. J Biol Chem. 281:37576–37585. 2006. View Article : Google Scholar

10 

Tang Y, Kesavan P, Nakada MT and Yan L: Tumor-stroma interaction: Positive feedback regulation of extracellular matrix metalloproteinase inducer (EMMPRIN) expression and matrix metalloproteinase-dependent generation of soluble EMMPRIN. Mol Cancer Res. 2:73–80. 2004.

11 

Taylor PM, Woodfield RJ, Hodgkin MN, Pettitt TR, Martin A, Kerr DJ and Wakelam MJ: Breast cancer cell-derived EMMPRIN stimulates fibroblast MMP2 release through a phospholipase A(2) and 5-lipoxygenase catalyzed pathway. Oncogene. 21:5765–5772. 2002. View Article : Google Scholar

12 

Yanaba K, Asano Y, Tada Y, Sugaya M, Kadono T, Hamaguchi Y and Sato S: Increased serum soluble CD147 levels in patients with systemic sclerosis: Association with scleroderma renal crisis. Clin Rheumatol. 31:835–839. 2012. View Article : Google Scholar

13 

Wu J, Hao ZW, Zhao YX, Yang XM, Tang H, Zhang X, Song F, Sun XX, Wang B, Nan G, et al: Full-length soluble CD147 promotes MMP-2 expression and is a potential serological marker in detection of hepatocellular carcinoma. J Transl Med. 12:1902014. View Article : Google Scholar :

14 

Hornbeck PV: Enzyme-linked immunosorbent assays. Curr Protoc Immunol. 110:1–23. 2015.

15 

Moonsom S, Tayapiwatana C, Wongkham S, Kongtawelert P and Kasinrerk W: A competitive ELISA for quantifying serum CD147: Reduction of soluble CD147 levels in cancer patient sera. Hybridoma (Larchmt). 29:45–52. 2010. View Article : Google Scholar

16 

Ku XM, Liao CG, Li Y, Yang XM, Yang B, Yao XY, Wang L, Kong LM, Zhao P and Chen ZN: Epitope mapping of series of monoclonal antibodies against the hepatocellular carcinoma-associated antigen HAb18G/CD147. Scand J Immunol. 65:435–443. 2007. View Article : Google Scholar

17 

Nelson PN, Westwood OM, Jefferis R, Goodall M and Hay FC: Characterisation of anti-IgG monoclonal antibody A57H by epitope mapping. Biochem Soc Trans. 25:373S1997. View Article : Google Scholar

18 

Lipman NS, Jackson LR, Trudel LJ and Weis-Garcia F: Monoclonal versus polyclonal antibodies: Distinguishing characteristics, applications, and information resources. ILAR J. 46:258–268. 2005. View Article : Google Scholar

19 

Lane D: Antibodies: A Laboratory Manual. Cold Spring Harbor Press; Cold Spring Harbor, N.Y: pp. 288–312. 1998

20 

Andrew SM and Titus JA: Purification of immunoglobulin G. Curr Protoc Immunol: Chapter. 2:Unit 2.72001.

21 

Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar

22 

Chen ZN and Liu YF: Monoclonal antibody HAb18 to human hepatoma. J Monocl Antibod. 3:15–17. 1990.(In Chinese).

23 

Howard GC and Bethell DR: Basic methods in antibody production and characterization. CRC Press Ltd; Boca Raton, Florida: pp. 1–49. 2000, View Article : Google Scholar

24 

Verdoliva A, Basile G and Fassina G: Affinity purification of immunoglobulins from chicken egg yolk using a new synthetic ligand. J Chromatogr B Biomed Sci Appl. 749:233–242. 2000. View Article : Google Scholar

25 

Hage DS: Survey of recent advances in analytical applications of immunoaffinity chromatography. J Chromatogr B Biomed Sci Appl. 715:3–28. 1998. View Article : Google Scholar

26 

Hage DS and Phillips TM: Immunoaffinity chromatographyTaylor & Francis, Handbook of Affinity Chromatography. New York: 62006

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Liu S, Li S, Zhang Y, Wang Y, Zhu Y, Wang B and Chen ZN: Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Mol Med Rep 15: 4035-4040, 2017.
APA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., & Chen, Z. (2017). Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Molecular Medicine Reports, 15, 4035-4040. https://doi.org/10.3892/mmr.2017.6523
MLA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15.6 (2017): 4035-4040.
Chicago
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15, no. 6 (2017): 4035-4040. https://doi.org/10.3892/mmr.2017.6523
Copy and paste a formatted citation
x
Spandidos Publications style
Liu S, Li S, Zhang Y, Wang Y, Zhu Y, Wang B and Chen ZN: Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Mol Med Rep 15: 4035-4040, 2017.
APA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., & Chen, Z. (2017). Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Molecular Medicine Reports, 15, 4035-4040. https://doi.org/10.3892/mmr.2017.6523
MLA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15.6 (2017): 4035-4040.
Chicago
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15, no. 6 (2017): 4035-4040. https://doi.org/10.3892/mmr.2017.6523
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team