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Purification of anti-bromelain antibodies by affinity precipitation using pNIPAm-linked bromelain.
Prep Biochem Biotechnol 2016; 46(2):161-4PB

Abstract

Affinity precipitation has emerged as a very useful technique for the purification of proteins. Here it has been employed for the purification of anti-bromelain antibodies from rabbit serum. A system has been developed for reversibly binding and thermoprecipitating antibodies. Anti-bromelain antibodies were raised in rabbit by immunizing it with bromelain. Poly-N-isopropylacrylamide (pNIPAm)-bromelain conjugate was prepared and incubated with rabbit serum. After that the temperature was raised for thermal precipitation of the polymer. Antibodies were then eluted from the complex by incubating it with a small volume of buffer, pH 3.0. This method is very effective in concentrating the antibodies. Purity and specificity of the antibodies were checked by gel electrophoresis and enzyme-linked immunosorbent assay (ELISA), respectively. The study of the effect of pH and temperature on the binding of the antibodies to the conjugate showed that the optimum binding occurred at pH 8.0 and 25°C.The polymer enzyme conjugate was further used for another cycle.

Authors+Show Affiliations

a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh , India.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

25569629

Citation

Mahmood, Rubab. "Purification of Anti-bromelain Antibodies By Affinity Precipitation Using pNIPAm-linked Bromelain." Preparative Biochemistry & Biotechnology, vol. 46, no. 2, 2016, pp. 161-4.
Mahmood R. Purification of anti-bromelain antibodies by affinity precipitation using pNIPAm-linked bromelain. Prep Biochem Biotechnol. 2016;46(2):161-4.
Mahmood, R. (2016). Purification of anti-bromelain antibodies by affinity precipitation using pNIPAm-linked bromelain. Preparative Biochemistry & Biotechnology, 46(2), pp. 161-4. doi:10.1080/10826068.2014.996232.
Mahmood R. Purification of Anti-bromelain Antibodies By Affinity Precipitation Using pNIPAm-linked Bromelain. Prep Biochem Biotechnol. 2016;46(2):161-4. PubMed PMID: 25569629.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification of anti-bromelain antibodies by affinity precipitation using pNIPAm-linked bromelain. A1 - Mahmood,Rubab, PY - 2015/1/9/entrez PY - 2015/1/9/pubmed PY - 2016/10/7/medline KW - Affinity precipitation KW - anti-bromelain antibodies KW - pNIPAm-bromelain KW - purification KW - thermosensitive polymer SP - 161 EP - 4 JF - Preparative biochemistry & biotechnology JO - Prep. Biochem. Biotechnol. VL - 46 IS - 2 N2 - Affinity precipitation has emerged as a very useful technique for the purification of proteins. Here it has been employed for the purification of anti-bromelain antibodies from rabbit serum. A system has been developed for reversibly binding and thermoprecipitating antibodies. Anti-bromelain antibodies were raised in rabbit by immunizing it with bromelain. Poly-N-isopropylacrylamide (pNIPAm)-bromelain conjugate was prepared and incubated with rabbit serum. After that the temperature was raised for thermal precipitation of the polymer. Antibodies were then eluted from the complex by incubating it with a small volume of buffer, pH 3.0. This method is very effective in concentrating the antibodies. Purity and specificity of the antibodies were checked by gel electrophoresis and enzyme-linked immunosorbent assay (ELISA), respectively. The study of the effect of pH and temperature on the binding of the antibodies to the conjugate showed that the optimum binding occurred at pH 8.0 and 25°C.The polymer enzyme conjugate was further used for another cycle. SN - 1532-2297 UR - https://www.unboundmedicine.com/medline/citation/25569629/Purification_of_anti_bromelain_antibodies_by_affinity_precipitation_using_pNIPAm_linked_bromelain_ L2 - http://www.tandfonline.com/doi/full/10.1080/10826068.2014.996232 DB - PRIME DP - Unbound Medicine ER -