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International Journal of Molecular Medicine
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December 2013 Volume 32 Issue 6

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Article

Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization

  • Authors:
    • Yan Wu
    • Desheng Zou
    • Yuhua Cao
    • Nannan Yao
    • Junye Wang
    • Wenhan Wang
    • Hongyu Jiang
    • Guiying Li
  • View Affiliations / Copyright

    Affiliations: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, College of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China, Department of General Internal Medicine, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
  • Pages: 1451-1457
    |
    Published online on: October 11, 2013
       https://doi.org/10.3892/ijmm.2013.1523
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Abstract

Cyclin D1 plays an important role in cell cycle progression. Increasing evidence indicates that cyclin D1 is overexpressed in the majority of tumor cells and has become a potential target for tumor therapy. However, little research has been done on the specific inhibition of cyclin D1 for cancer therapy. With the rapid development of the phage display antibody library technique, single-chain variable fragment (scFv) antibodies have emerged, which have tremendous application prospects in cancer therapy and diagonosis. In this study, a human scFv binding specifically to cyclin D1 (AD5) that was derived from a human semi-synthetic scFv phage library was expressed in the soluble form in Escherichia coli (E. coli) HB2151 cells. To characterize AD5, soluble AD5 was purified successfully through ammonium sulfate precipitation and affinity chromatography from the culture supernatant of AD5/HB2151. ELISA assay revealed that purified soluble AD5 could specifically bind to human recombinant cyclin D1 with approximately (1.19±0.056) x 107 M-1 affinity constant and showed approximately 52% competitive inhibition with the anti-cyclin D1 polyclonal antibody for binding to cyclin D1 in vitro. These results suggest that the scFv antibody against cyclin D1 may be a novel potential tool for targeting cyclin D1 in cancer therapy and diagnosis.
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1 

Sherr CJ: Mammalian G1 cyclins. Cell. 73:1059–1065. 1993. View Article : Google Scholar : PubMed/NCBI

2 

Malumbres M and Barbacid M: Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 9:153–166. 2009. View Article : Google Scholar : PubMed/NCBI

3 

Deshpande A, Sicinski P and Hinds PW: Cyclins and CDKs in development and cancer: a perspective. Oncogene. 24:2909–2915. 2005. View Article : Google Scholar : PubMed/NCBI

4 

Fu M, Wang C, Li Z, Sakamaki T and Pestell RG: Minireview: cyclin D1: normal and abnormal functions. Endocrinology. 145:5439–5447. 2004. View Article : Google Scholar : PubMed/NCBI

5 

Musgrove EA, Caldon CE, Barraclough J, Stone A and Sutherland RL: Cyclin D as a therapeutic target in cancer. Nat Rev Cancer. 11:558–572. 2011. View Article : Google Scholar : PubMed/NCBI

6 

Roy PG, Pratt N, Purdie CA, Baker L, Ashfield A, Quinlan P and Thompson AM: High CCND1 amplification identifies a group of poor prognosis women with estrogen receptor positive breast cancer. Int J Cancer. 127:355–360. 2010.PubMed/NCBI

7 

Yu Q, Geng Y and Sicinski P: Specific protection against breast cancers by cyclin D1 ablation. Nature. 411:1017–1021. 2001. View Article : Google Scholar : PubMed/NCBI

8 

Choi YJ, Li X, Hydbring P, Sanda T, Stefano J, Christie AL, Signoretti S, Look AT, Kung AL, von Boehmer H and Sicinski P: The requirement for cyclin D function in tumor maintenance. Cancer Cell. 22:438–451. 2012. View Article : Google Scholar : PubMed/NCBI

9 

Malumbres M: Cell cycle-based therapies move forward. Cancer Cell. 22:419–420. 2012. View Article : Google Scholar : PubMed/NCBI

10 

Beroukhim R, Mermel CH, Porter D, et al: The landscape of somatic copy-number alteration across human cancers. Nature. 463:899–905. 2010. View Article : Google Scholar : PubMed/NCBI

11 

Kim JK and Diehl JA: Nuclear cyclin D1: an oncogenic driver in human cancer. J Cell Physiol. 220:292–306. 2009. View Article : Google Scholar : PubMed/NCBI

12 

Scott AM, Wolchok JD and Old LJ: Antibody therapy of cancer. Nat Rev Cancer. 12:278–287. 2012. View Article : Google Scholar

13 

Elvin JG, Couston RG and van der Walle CF: Therapeutic antibodies: market considerations, disease targets and bioprocessing. Int J Pharm. 440:83–98. 2013. View Article : Google Scholar : PubMed/NCBI

14 

Stewart CS, MacKenzie CR and Hall JC: Isolation, characterization and pentamerization of alpha-cobrotoxin specific single-domain antibodies from a naïve phage display library: preliminary findings for antivenom development. Toxicon. 49:699–709. 2007.PubMed/NCBI

15 

Wen S, Zhang X, Liu Y, Zhang QQ, Liu XJ and Liang JS: Selection of a single chain variable fragment antibody against ivermectin from a phage displayed library. J Agric Food Chem. 58:5387–5391. 2010. View Article : Google Scholar : PubMed/NCBI

16 

Holliger P and Hudson PJ: Engineered antibody fragments and the rise of single domains. Nat Biotechnol. 23:1126–1136. 2005. View Article : Google Scholar : PubMed/NCBI

17 

Dai H, Gao H, Zhao X, Dai L, Zhang X, Xiao N, Zhao R and Hemmingsen SM: Construction and characterization of a novel recombinant single-chain variable fragment antibody against white spot syndrome virus from shrimp. J Immunol Methods. 279:267–275. 2003. View Article : Google Scholar : PubMed/NCBI

18 

Weisser NE and Hall JC: Applications of single-chain variable fragment antibodies in therapeutics and diagnostics. Biotechnol Adv. 27:502–520. 2009. View Article : Google Scholar : PubMed/NCBI

19 

Zhou LH, Zhu X, Cao YH, Wang L, Chen Y, Du BR and Li GY: Construction of expression vector for anti-cyclin D1 intrabody AD5N and its inhibitory effects on cell proliferation of breast cancer. Chin J Immunol. 24:703–706. 2008.

20 

Zhou LH, Zhu X, Cao YH, Chen Y, Tian Y, Wang L and Li GY: Effects of anti-cyclin D1 intrabody AD5N on HeLa cells of uterine cervix cancer. Chin J Clin Oncol. 35:942–944. 2008.

21 

Qiao YY, Wang Y, Chen XS, Wang YX and Hua B: Construction of a large single-chain phage antibody library. Chin J Microbiol Immunol. 24:194–197. 2004.

22 

Li GY, Zou DS and Zhou LH: Expression and purification of recombinant human cyclin D1 in E. coli BL21. J Jilin Univ. 44:839–843. 2006.

23 

Beatty JD, Beatty BG and Vlahos WG: Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Methods. 100:173–179. 1987. View Article : Google Scholar : PubMed/NCBI

24 

Ding Z, Wu CJ, Chu GC, et al: SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression. Nature. 470:269–273. 2011. View Article : Google Scholar : PubMed/NCBI

25 

Bienvenu F, Jirawatnotai S, Elias JE, et al: Transcriptional role of cyclin D1 in development revealed by a genetic-proteomic screen. Nature. 463:374–378. 2010. View Article : Google Scholar : PubMed/NCBI

26 

Bartek J and Lukas J: DNA repair: Cyclin D1 multitasks. Nature. 474:171–172. 2011. View Article : Google Scholar : PubMed/NCBI

27 

Ahmad ZA, Yeap SK, Ali AM, Ho WY, Alitheen NB and Hamid M: scFv antibody: principles and clinical application. Clin Dev Immunol. 2012:9802502012. View Article : Google Scholar : PubMed/NCBI

28 

Zhu X, Wang D, Li S, Xiao Q, Tao K, Hu J, Cao W and Feng W: Expression of a humanized single-chain variable fragment antibody targeting chronic myeloid leukemia cells in Escherichia coli and its characterization. Int J Mol Med. 29:939–945. 2012.PubMed/NCBI

29 

Sushma K, Vijayalakshmi MA, Krishnan V and Satheeshkumar PK: Cloning, expression, purification and characterization of a single chain variable fragment specific to tumor necrosis factor alpha in Escherichia coli. J Biotechnol. 156:238–244. 2010. View Article : Google Scholar : PubMed/NCBI

30 

Yang T, Yang L, Chai W, Li R, Xie J and Niu B: A strategy for high-level expression of a single-chain variable fragment against TNFα by subcloning antibody variable regions from the phage display vector pCANTAB 5E into pBV220. Protein Expr Purif. 76:109–114. 2011.PubMed/NCBI

31 

Abdolalizadeh J, Nouri M, Zolbanin JM, Baradaran B, Barzegari A and Omidi Y: Downstream characterization of anti-TNF-α single chain variable fragment antibodies. Hum Antibodies. 21:41–48. 2012.

32 

Adams GP and Schier R: Generating improved single-chain Fv molecules for tumor targeting. J Immunol Methods. 231:249–260. 1999. View Article : Google Scholar : PubMed/NCBI

33 

Yuan W and Parrish CR: Comparison of two single-chain antibodies that neutralize canine parovirus: analysis of an antibody-combining site and mechanisms of neutralization. Virology. 269:471–480. 2000. View Article : Google Scholar : PubMed/NCBI

34 

Neri P, Shigemori N, Hamada-Tsutsumi S, Tsukamoto K, Arimitsu H, Shimizu T, Akahori Y, Kurosawa Y and Tsuji T: Single chain variable fragment antibodies against Shiga toxins isolated from a human antibody phage display library. Vaccine. 29:5340–5346. 2011. View Article : Google Scholar : PubMed/NCBI

35 

Schier R, McCall A, Adams GP, Marshall KW, Merritt H, Yim M, Crawford RS, Weiner LM, Marks C and Marks JD: Isolation of picomolar affinity anti-c-erbB-2 single-chain Fv by molecular evolution of the complementarity determining regions in the center of the antibody binding site. J Mol Biol. 263:551–567. 1996. View Article : Google Scholar

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Copy and paste a formatted citation
Spandidos Publications style
Wu Y, Zou D, Cao Y, Yao N, Wang J, Wang W, Jiang H and Li G: Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization. Int J Mol Med 32: 1451-1457, 2013.
APA
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W. ... Li, G. (2013). Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization. International Journal of Molecular Medicine, 32, 1451-1457. https://doi.org/10.3892/ijmm.2013.1523
MLA
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W., Jiang, H., Li, G."Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization". International Journal of Molecular Medicine 32.6 (2013): 1451-1457.
Chicago
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W., Jiang, H., Li, G."Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization". International Journal of Molecular Medicine 32, no. 6 (2013): 1451-1457. https://doi.org/10.3892/ijmm.2013.1523
Copy and paste a formatted citation
x
Spandidos Publications style
Wu Y, Zou D, Cao Y, Yao N, Wang J, Wang W, Jiang H and Li G: Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization. Int J Mol Med 32: 1451-1457, 2013.
APA
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W. ... Li, G. (2013). Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization. International Journal of Molecular Medicine, 32, 1451-1457. https://doi.org/10.3892/ijmm.2013.1523
MLA
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W., Jiang, H., Li, G."Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization". International Journal of Molecular Medicine 32.6 (2013): 1451-1457.
Chicago
Wu, Y., Zou, D., Cao, Y., Yao, N., Wang, J., Wang, W., Jiang, H., Li, G."Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization". International Journal of Molecular Medicine 32, no. 6 (2013): 1451-1457. https://doi.org/10.3892/ijmm.2013.1523
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