Tags

Type your tag names separated by a space and hit enter

Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice.
Di Yi Jun Yi Da Xue Xue Bao. 2003 Jul; 23(7):640-2.DY

Abstract

OBJECTIVE

To develop a rapid and efficient method for preparing monoclonal antibodies (mAb) against SARS-associated coronavirus (SARS-Cov) nucleocapsid (N) protein.

METHODS

BALB/c mice were injected with the recombinant N protein of SARS-Cov into the foot-pads for the immunization, and the popliteal lymph nodes were isolated 15 d later for mAb-producing hybridomas, from which the mAbs against the N protein of SARS-Cov were screened. The identification of the mAb against the N protein of SARS-Cov was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting.

RESULTS

Four strains of hybridomas were obtained that produced the mAb specific to the N protein without detectable cross-reactivity with other pathogens. Of the 4 strains, 2 were identified as the immunoglobulin G1 (IgG1) isotype, 1 IgG2a, and the other IgG2b, with affinity constants (Ka) of 2 of the strains being 4.14 x 10(-9)M and 3.19 x 10(-9)M respectively.

CONCLUSION

This is the first report on the preparation of mAb that is specific to the SARS-Cov, and the high-specificity and high-affinity mAb produced by the 4 strains of hybridomas provide a basis for further researches on the pathogenesis and early diagnosis of SARS.

Authors+Show Affiliations

Central Laboratory, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China. linche@pub.guangzhou.gd.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12865208

Citation

Che, Xiao-yan, et al. "Rapid and Efficient Preparation of Monoclonal Antibodies Against SARS-associated Coronavirus Nucleocapsid Protein By Immunizing Mice." Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA, vol. 23, no. 7, 2003, pp. 640-2.
Che XY, Qiu LW, Pan YX, et al. Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice. Di Yi Jun Yi Da Xue Xue Bao. 2003;23(7):640-2.
Che, X. Y., Qiu, L. W., Pan, Y. X., Xu, H., Hao, W., Liao, Z. Y., Mei, Y. B., Zhang, L. Y., Wan, Z. Y., Yuan, G. Y., & Huang, Z. (2003). Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice. Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA, 23(7), 640-2.
Che XY, et al. Rapid and Efficient Preparation of Monoclonal Antibodies Against SARS-associated Coronavirus Nucleocapsid Protein By Immunizing Mice. Di Yi Jun Yi Da Xue Xue Bao. 2003;23(7):640-2. PubMed PMID: 12865208.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice. AU - Che,Xiao-yan, AU - Qiu,Li-wen, AU - Pan,Yu-xian, AU - Xu,Hua, AU - Hao,Wei, AU - Liao,Zhi-yong, AU - Mei,Ya-bo, AU - Zhang,Li-ya, AU - Wan,Zhuo-yue, AU - Yuan,Guo-yong, AU - Huang,Zhen, PY - 2003/7/17/pubmed PY - 2004/1/30/medline PY - 2003/7/17/entrez SP - 640 EP - 2 JF - Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA JO - Di Yi Jun Yi Da Xue Xue Bao VL - 23 IS - 7 N2 - OBJECTIVE: To develop a rapid and efficient method for preparing monoclonal antibodies (mAb) against SARS-associated coronavirus (SARS-Cov) nucleocapsid (N) protein. METHODS: BALB/c mice were injected with the recombinant N protein of SARS-Cov into the foot-pads for the immunization, and the popliteal lymph nodes were isolated 15 d later for mAb-producing hybridomas, from which the mAbs against the N protein of SARS-Cov were screened. The identification of the mAb against the N protein of SARS-Cov was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. RESULTS: Four strains of hybridomas were obtained that produced the mAb specific to the N protein without detectable cross-reactivity with other pathogens. Of the 4 strains, 2 were identified as the immunoglobulin G1 (IgG1) isotype, 1 IgG2a, and the other IgG2b, with affinity constants (Ka) of 2 of the strains being 4.14 x 10(-9)M and 3.19 x 10(-9)M respectively. CONCLUSION: This is the first report on the preparation of mAb that is specific to the SARS-Cov, and the high-specificity and high-affinity mAb produced by the 4 strains of hybridomas provide a basis for further researches on the pathogenesis and early diagnosis of SARS. SN - 1000-2588 UR - https://www.unboundmedicine.com/medline/citation/12865208/Rapid_and_efficient_preparation_of_monoclonal_antibodies_against_SARS_associated_coronavirus_nucleocapsid_protein_by_immunizing_mice_ L2 - https://www.lens.org/lens/search/patent/list?q=citation_id:12865208 DB - PRIME DP - Unbound Medicine ER -