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
February-2020 Volume 21 Issue 2

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
February-2020 Volume 21 Issue 2

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

  • Supplementary Files
    • Supplementary_Data.pdf
Article Open Access

Affinity improvement of the fully human anti‑TSLP recombinant antibody

  • Authors:
    • Qi Chen
    • Dequn Xian
    • Wenfeng Xu
    • Siji Nian
    • Hong Yu
    • Yuchuan Wu
    • Qing Yuan
  • View Affiliations / Copyright

    Affiliations: State Key Laboratory of Quality Research in Chinese Medicines, Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Taipa, Macau SAR 000853, P.R. China, Immunology Department, The School of Basic Medical Sciences, Southwest Medical University, Luzhou, Sichuan 646000, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 759-767
    |
    Published online on: December 12, 2019
       https://doi.org/10.3892/mmr.2019.10880
  • 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

Thymic stromal lymphopoietin (TSLP) is a potentially important target for the treatment of asthma and malignancies. However, a fully human antibody reactive with TSLP is currently unavailable for clinical use. In a previous study, a human anti‑TSLP‑single‑chain antibody variable fragment (anti‑TSLP‑scFv) 84 was selected by phage display from a constructed human scFv library. In the present study, a computer simulation method was developed using Discovery Studio 4.5 software, to increase the affinity of anti‑TSLP‑scFv‑84. Specific primers were designed and mutated DNA sequences of anti‑TSLP‑scFvs were obtained by overlap extension PCR. The mutant scFvs were expressed in pLZ16 and affinity‑enhanced anti‑TSLP‑scFv‑M4 was screened using ELISA. However, in general the scFvs have low stability and short half‑lives in vivo. Therefore, scFv‑84 and scFv‑M4 were inserted into eukaryotic expression vectors (pcDNA3.1‑sp‑Fc and PMH3EN‑sp‑Fc) and then transfected into 293F cells to express anti‑TSLP‑scFv‑Fc. ELISA and western blotting results indicated the size of the anti‑TSLP‑scFv‑Fc to be ~50 kDa. Binding of anti‑TSLP‑scFv‑Fc‑M4 to TSLP was enhanced compared with the pre‑mutated scFv‑Fc‑84. The affinity of the mutated recombinant antibody was determined using the BIAcore technique and found to be ~10‑fold greater than the pre‑mutated antibody.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

View References

1 

Friend SL, Hosier S, Nelson A, Foxworthe D, Williams DE and Farr A: A thymic stromal cell line supports in vitro development of surface IgM+ B cells and produces a novel growth factor affecting B and T lineage cells. Exp Hematol. 22:321–328. 1994.PubMed/NCBI

2 

Tatsuno K, Fujiyama T, Yamaguchi H, Waki M and Tokura Y: TSLP directly interacts with skin-homing Th2 cells highly expressing its receptor to enhance IL-4 production in atopic dermatitis. J Invest Dermatol. 135:3017–3024. 2015. View Article : Google Scholar : PubMed/NCBI

3 

Park JH, Jeong DY, Peyrin-Biroulet L, Eisenhut M and Shin JI: Insight into the role of TSLP in inflammatory bowel diseases. Autoimmun Rev. 16:55–63. 2017. View Article : Google Scholar : PubMed/NCBI

4 

Kuan EL and Ziegler SF: A tumor-myeloid cell axis, mediated via the cytokines IL-1α and TSLP, promotes the progression of breast cancer. Nat Immunol. 19:366–374. 2018. View Article : Google Scholar : PubMed/NCBI

5 

Manoutcharian K, Perez-Garmendia R and Gevorkian G: Recombinant antibody fragments for neurodegenerative diseases. Curr Neuropharmacol. 15:779–788. 2017. View Article : Google Scholar : PubMed/NCBI

6 

Shehata L, Maurer DP, Wec AZ, Lilov A, Champney E, Sun T, Archambault K, Burnina I, Lynaugh H, Zhi X, et al: Affinity maturation enhances antibody specificity but compromises conformational stability. Cell Rep. 28:3300–3308.e4. 2019. View Article : Google Scholar : PubMed/NCBI

7 

Nakamura T, Motoyama T, Hirokawa T, Hirono S and Yamaguchi I: Computer-aided modeling of pentachlorophenol 4-monooxygenase and site-directed mutagenesis of its active site. Chem Pharm Bull (Tokyo). 51:1293–1298. 2003. View Article : Google Scholar : PubMed/NCBI

8 

Chen JM, Grad R, Monaco R and Pincus MR: Prediction of the three-dimensional structure of the rap-1A protein from its homology to the ras-gene-encoded p21 protein. J Protein Chem. 15:11–15. 1996. View Article : Google Scholar : PubMed/NCBI

9 

Yuan Q, Huang L, Wang X, Wu Y, Gao Y, Li C and Nian S: Construction of human nonimmune library and selection of scFvs against IL-33. Appl Biochem Biotechnol. 167:498–509. 2012. View Article : Google Scholar : PubMed/NCBI

10 

Sircar A: Methods for the homology modeling of antibody variable regions. Methods Mol Biol. 857:301–311. 2012. View Article : Google Scholar : PubMed/NCBI

11 

Nian S, Wu T, Ye Y, Wang X, Xu W and Yuan Q: Development and identification of fully human scFv-Fcs against Staphylococcus aureus. BMC Immunol. 17:82016. View Article : Google Scholar : PubMed/NCBI

12 

Ye Y, Nian S, Xu W, Wu T, Wang X, Gao Y and Yuan Q: Construction and expression of human scFv-Fc against interleukin-33. Protein Expr Purif. 114:58–63. 2015. View Article : Google Scholar : PubMed/NCBI

13 

Nigi I, Fairall L and Schwabe JWR: Expression and purification of protein complexes suitable for structural studies using mammalian HEK 293F cells. Curr Protoc Protein Sci. 90:5.28.1–5.28.16. 2017. View Article : Google Scholar

14 

Portolano N, Watson PJ, Fairall L, Millard CJ, Milano CP, Song Y, Cowley SM and Schwabe JW: Recombinant protein expression for structural biology in HEK 293F suspension cells: A novel and accessible approach. J Vis Exp. e518972014.PubMed/NCBI

15 

Molek P, Vodnik M, Strukelj B and Bratkovič T: Screening of synthetic phage display scFv libraries yields competitive ligands of human leptin receptor. Biochem Biophys Res Commun. 452:479–483. 2014. View Article : Google Scholar : PubMed/NCBI

16 

Yuan Q, Wang Z, Nian S, Yin Y, Chen G and Xia Y: Screening of high-affinity scFvs from a ribosome displayed library using BIAcore biosensor. Appl Biochem Biotechnol. 152:224–234. 2009. View Article : Google Scholar : PubMed/NCBI

17 

Kahramanoğlu Aksoy E, Akpınar MY, Pirinççi Sapmaz F, Doğan Ö, Uzman M and Nazlıgül Y: Thymic stromal lymphopoietin levels are increased in patients with celiac disease. Bosn J Basic Med Sci. 19:282–287. 2019.PubMed/NCBI

18 

Tahaghoghi-Hajghorbani S, Ajami A, Ghorbanalipoor S, Hosseini-Khah Z, Taghiloo S, Khaje-Enayati P and Hosseini V: Protective effect of TSLP and IL-33 cytokines in ulcerative colitis. Auto Immun Highlights. 10:12019. View Article : Google Scholar : PubMed/NCBI

19 

Borowski A, Vetter T, Kuepper M, Wohlmann A, Krause S, Lorenzen T, Virchow JC, Luttmann W and Friedrich K: Expression analysis and specific blockade of the receptor for human thymic stromal lymphopoietin (TSLP) by novel antibodies to the human TSLPRα receptor chain. Cytokine. 61:546–555. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Yazdi IK, Taghipour N, Hmaidan S, Palomba R, Scaria S, Munoz A, Boone TB and Tasciotti E: Antibody-mediated inhibition of Nogo-A signaling promotes neurite growth in PC-12 cells. J Tissue Eng. 7:20417314166297672016. View Article : Google Scholar : PubMed/NCBI

21 

Hafeez U, Gan HK and Scott AM: Monoclonal antibodies as immunomodulatory therapy against cancer and autoimmune diseases. Curr Opin Pharmacol. 41:114–121. 2018. View Article : Google Scholar : PubMed/NCBI

22 

Moutel S, Bery N, Bernard V, Keller L, Lemesre E, de Marco A, Ligat L, Rain JC, Favre G, Olichon A and Perez F: NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies. Elife. 5:e162282016. View Article : Google Scholar : PubMed/NCBI

23 

Townsend S, Fennell BJ, Apgar JR, Lambert M, McDonnell B, Grant J, Wade J, Franklin E, Foy N, Ní Shúilleabháin D, et al: Augmented binary substitution: Single-pass CDR germ-lining and stabilization of therapeutic antibodies. Proc Natl Acad Sci USA. 112:15354–15359. 2015. View Article : Google Scholar : PubMed/NCBI

24 

Rodgers KR and Chou RC: Therapeutic monoclonal antibodies and derivatives: Historical perspectives and future directions. Biotechnol Adv. 34:1149–1158. 2016. View Article : Google Scholar : PubMed/NCBI

25 

Nian S, Zhu J, Yu H, Chen Q, Ye Y, Cao X and Yuan Q: Development and identification of a fully human single-chain variable fragment 29 against TSLP. Biotechnol Appl Biochem. 66:510–516. 2019. View Article : Google Scholar : PubMed/NCBI

26 

Zahnd C, Spinelli S, Luginbühl B, Amstutz P, Cambillau C and Plückthun A: Directed in vitro evolution and crystallographic analysis of a peptide-binding single chain antibody fragment (scFv) with low picomolar affinity. J Biol Chem. 279:18870–18877. 2004. View Article : Google Scholar : PubMed/NCBI

27 

Lewis CM, Hollis GF, Mark GE III, Tung JS and Ludmerer SW: Use of a novel mutagenesis strategy, optimized residue substitution, to decrease the off-rate of an anti-gp120 antibody. Mol Immunol. 32:1065–1072. 1995. View Article : Google Scholar : PubMed/NCBI

28 

Tiller KE, Chowdhury R, Li T, Ludwig SD, Sen S, Maranas CD and Tessier PM: Facile affinity maturation of antibody variable domains using natural diversity mutagenesis. Front Immunol. 8:9862017. View Article : Google Scholar : PubMed/NCBI

29 

Zhang YF and Ho M: Humanization of high-affinity antibodies targeting glypican-3 in hepatocellular carcinoma. Sci Rep. 6:338782016. View Article : Google Scholar : PubMed/NCBI

30 

Ahmed M, Cheng M, Zhao Q, Goldgur Y, Cheal SM, Guo HF, Larson SM and Cheung NK: Humanized affinity-matured monoclonal antibody 8H9 has potent antitumor activity and binds to FG loop of tumor antigen B7-H3. J Biol Chem. 290:30018–30029. 2015. View Article : Google Scholar : PubMed/NCBI

31 

Subedi GP, Johnson RW, Moniz HA, Moremen KW and Barb A: High yield expression of recombinant human protein with the transient transfection of HEK293 cells in suspension. J Vis Exp. e535682015.PubMed/NCBI

32 

Mitchell PD, El-Gammal AI and O'Byrne PM: Emerging monoclonal antibodies as targeted innovative therapeutic approaches to asthma. Clin Pharmacol Ther. 99:38–48. 2016. View Article : Google Scholar : PubMed/NCBI

33 

Cohan SL, Lucassen EB, Romba MC and Linch SN: Daclizumab: Mechanisms of action, therapeutic efficacy, adverse events and its uncovering the potential role of innate immune system recruitment as a treatment strategy for relapsing multiple sclerosis. Biomedicines. 7:E182019. View Article : Google Scholar : PubMed/NCBI

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Chen Q, Xian D, Xu W, Nian S, Yu H, Wu Y and Yuan Q: Affinity improvement of the fully human anti‑TSLP recombinant antibody. Mol Med Rep 21: 759-767, 2020.
APA
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., & Yuan, Q. (2020). Affinity improvement of the fully human anti‑TSLP recombinant antibody. Molecular Medicine Reports, 21, 759-767. https://doi.org/10.3892/mmr.2019.10880
MLA
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., Yuan, Q."Affinity improvement of the fully human anti‑TSLP recombinant antibody". Molecular Medicine Reports 21.2 (2020): 759-767.
Chicago
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., Yuan, Q."Affinity improvement of the fully human anti‑TSLP recombinant antibody". Molecular Medicine Reports 21, no. 2 (2020): 759-767. https://doi.org/10.3892/mmr.2019.10880
Copy and paste a formatted citation
x
Spandidos Publications style
Chen Q, Xian D, Xu W, Nian S, Yu H, Wu Y and Yuan Q: Affinity improvement of the fully human anti‑TSLP recombinant antibody. Mol Med Rep 21: 759-767, 2020.
APA
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., & Yuan, Q. (2020). Affinity improvement of the fully human anti‑TSLP recombinant antibody. Molecular Medicine Reports, 21, 759-767. https://doi.org/10.3892/mmr.2019.10880
MLA
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., Yuan, Q."Affinity improvement of the fully human anti‑TSLP recombinant antibody". Molecular Medicine Reports 21.2 (2020): 759-767.
Chicago
Chen, Q., Xian, D., Xu, W., Nian, S., Yu, H., Wu, Y., Yuan, Q."Affinity improvement of the fully human anti‑TSLP recombinant antibody". Molecular Medicine Reports 21, no. 2 (2020): 759-767. https://doi.org/10.3892/mmr.2019.10880
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