JBM  Vol.3 No.3 , March 2015
A Preliminary Study on the Preparation of Monoclonal Antibody with Umbilical Cord Blood Cell
ABSTRACT

Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, then incubated them and collected cell supernatant. By using Caprylic acid- saturated Ammonium sulfate method, monoclonal antibody was purified, and IgG subtype identification was conducted by their subtype classification kit of sigma. In traditional way, the preparation of monoclonal antibody cannot do without BABL/C mice and SP2/0 hybridoma cells. Consider this kinds of monoclonal antibody as a positive control, the reactivity and specificity of monoclonal antibodies were identified by Dot-ELISA. Results: By ELISA, we obtained four strains of positive umbilical cord blood cells. After subculture, cryopreservation and resuscitation in vitro, three of them were confirmed to secrete monoclonal antibody against rubella virus stably. The result of monoclonal antibody subtype classification showed that, both IgG1 and IgG2 types were detected. However, the quantity of monoclonal antibody prepared by umbilical cord blood cells was less than that from traditional method. Conclusions: The method of prepare monoclonal antibody using umbilical cord blood cells is feasible. 


Cite this paper
Su, D. , Zhang, S. , Zhang, S. , Gong, Y. , Zhu, S. , Zhang, L. , Wang, Y. , Yue, C. and Liu, X. (2015) A Preliminary Study on the Preparation of Monoclonal Antibody with Umbilical Cord Blood Cell. Journal of Biosciences and Medicines, 3, 73-77. doi: 10.4236/jbm.2015.33011.
References
[1]   Qian, Z.M., Guo, T.H., Wang, W., Guo, S.Y. and Yuan, Q.M. (2002) The Basic Technology and Clinical Application Research Status of Umbilical Cord Blood. Chinese Journal of Medicine, 41, 568-570.

[2]   Rubinstein, P., Carrier, C., Scara-davou, A., Kurtzberg, J., Adamson, J., Migliaccio, A.R., et al. (1998) Outcomes among 562 Recipients of Placental-Blood Transplants from Unrelated Donors. New England Journal of Medicine, 339, 1565-1577. http://dx.doi.org/10.1056/NEJM199811263392201

[3]   Gabutti, V., Timeus, F. and Ramenhi, U. (1993) Expansion of Cord Blood Progenitors and Use for Hemopoietic Reconstitution. Stem Cells Dayton, 11, 105-201. http://dx.doi.org/10.1002/stem.5530110818

[4]   Chen, J., Sanberg, P.R., Li, Y., Wang, L., Lu, M., Willing, A.E., et al. (2001) Intravenous Administration of Human Umbilical Cord Blood Reduces Behavioral Deficits after Stroke in Rats. Stroke, 32, 2682-2688. http://dx.doi.org/10.1161/hs1101.098367

[5]   Sanchez-Ramos, J.R., Song, S., Kamath, S.G., Zigova, T., Willing, A., Car-dozo-Pelaez, F., et al. (2001) Expression of Neural Markers in Human Umbilical Cord Blood. Experimental Neurology, 171, 109-115. http://dx.doi.org/10.1006/exnr.2001.7748

[6]   Ende, N., Chen, R. and Ende Harris, D. (2001) Human Umbilical Cord Blood Cells Ameliorate Alzheimer’s Disease in Transgenic Mice. A Brief Report. Journal of Medicine, 32, 241-247.

[7]   Ende, N. and Chen, R. (2001) Human Umbilical Cord Blood Cells Ameliorate Huntington’s Disease in Transgenic Mice. A Brief Report. Journal of Medicine, 231-240.

[8]   Wang, L.L. and Xu, H.X. (2013) Clinical Immunology Test. 5th Edition, People’s Medical Publishing House, Beijing, 2-11.

[9]   Jin, B.Q. (2008) Medical Immunology. 5th Edition, People’s Medical Publishing House, Beijing.

[10]   Zhu, H.P., Sun, Z.M., Wang, J., He, X.D., Dai, H.M., Wu, J.S., et al. (2002) Research on Biological Characteristic of Cord Blood Cell. Journal of Clinical Transfusion and Laboratory Medicine, 2, 8-10.

[11]   Liu, G.J., Guo, L.R. and Qiao, Z.H. (2002) Study on Biological Characteristic of Umbilical Cord Blood Cells. Shanxi Clinical Medicine Journal, 11, 406-410.

[12]   Huang, L.P., Liu, C.M., Wei, Y.W., Zhang, Z.X., Lu, Y.H. and Guo, L.J. (2009) Preparation and Identification of Neutralizating Monoclonal Antibody to the Recombinant Capsid Protein of Porcine Circovirus Type 2. Chinese Journal of Preventive Veterinary Medicine, 31, 132-136.

[13]   Chen, D., Sun, G.R. and Liu, Z.S. (2007) Application of Caprylic Acid Ammonium Sulfate Precipitation Method Incombination of Monoclonal Antibody Purification. Journal of Anhui Agricultural Sciences, 35, 8105-8108.

[14]   Zhou, Y., Li, Y.S., Pan, F.G., Tan, J.H., Liu, Z.S. and Wang, Z. (2006) The Study of Purified Methods of Mice Ascites IgG McAb. Heilongjiang Animal Science and Veterinary Medicine, 10, 14-16.

[15]   Dong, G.H., Wei, P., Huang, J.Q., Du, Q.B., Yue, H. and Du, W.S. (2014) IL-31 Establishment of ELISA Method for Detection of IL-31 and Preparation of the Monoclonal Antibody. Chinese Journal of Immunology, 4, 495-498.

[16]   Ti, J.F., Li, Z.J. and Li, F. (2012) The Isolation and Identification of Duck Hepatitis Virus I of Shandong Area and Development of Its Monoclonal Antibody. Progress in Veterinary Medicine, 33, 63-66.

[17]   Pan, P.P., Wen, L.Z., Shan, Z.Z., Ding, Y., Yuan, B. and Ren, W.S. (2011) Monoclonal Antibody against Duck Hepatitis Virus Preparation. Chinese Agricultural Science Bulletin, 27, 415-441.

[18]   Wang, W., Wang, X.M., Wen, L.B., He, K.W., Zhou, J.M., Guo, R.L., et al. (2014) Preparation and Identification of Specific Monoclonal Antibody against Porcine Circovirus Type 2. Agricultural Science and Technology, 15, 173-176.

[19]   Zhang, L., Wang, Y.Q., Hao, L.S., Gao, M.M., Li, A.D. and Tao, Y. (2014) Preparation of the Monoclonal Antibody against Cap Protein of Porcine Circovrius typ2 and Its Antigenic Epitope Identification. Chinese Journal of Preventive Veterinary Medicine, 4, 310-313.

[20]   Qi, X. (1999) The Preparation of the Monoclonal Antibody of Rubella Virus. Journal of Lanzhou Medical College, 25, 10-11.

 
 
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