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Production of double antibody for radioimmunoassay (sheep anti-rabbit IgG antiserum); Producao de duplo anticorpo para radioimunoensaio (antissoro de carneiro anti-IgG de coelho)

Abstract

A second antibody (sheep anti-rabbit IgG antiserum) to be used in RIAs in which the first antibody is raised in rabbits was produced. For this production, initially the IgG was isolated from rabbit serum and purified by sodium sulphate precipitation followed by ion exchange chromatography on DEAE-cellulose. Four sheep were immunized with 500 u g of purified rabbit IgG, emulsified in Freund Complete Adjuvant and administered by multisite subcutaneous injections. These injections were repeated at 20-days intervals and blood samples (40 ml) were taken from the jugular vein 10 days after the boosts for the evaluation of the antisera title. After each four boosts a great bleeding was done by the same route. Approximately 500 ml of serum were obtained in each bleeding per animal. The antisera were evaluated by the human thyrotropin RIA developed at IPEN laboratories employing reagents provided by NIDDKD, USA. These evaluations referred to the determination of the antisera title and of the ideal concentration of carrier IgG, to the study of the kinetic of precipitation and to the confirmation of the inexistent cross-reactivity with human IgG, in comparison with a reference antiserum of know precipitation characteristics supplied by the Radioassay System Laboratories. Approximately 3,6 l  More>>
Authors:
Publication Date:
Dec 31, 1993
Product Type:
Miscellaneous
Report Number:
INIS-BR-3486
Reference Number:
SCA: 553006; 550601; PA: AIX-26:040665; EDB-95:073634; ERA-20:016616; NTS-95:021965; SN: 95001381867
Resource Relation:
Other Information: TH: Tese (M.Sc.).; PBD: 1993
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; IMMUNE SERUMS; RADIOIMMUNOASSAY; SHEEP; IMMUNOGLOBULINS; RABBITS; DIAGRAMS; ELECTROPHORESIS; EXPERIMENTAL DATA; ION EXCHANGE CHROMATOGRAPHY; PRECIPITATION; PRODUCTION; PURIFICATION; SODIUM SULFATES; STORAGE
OSTI ID:
38160
Research Organizations:
Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)
Country of Origin:
Brazil
Language:
Portuguese
Other Identifying Numbers:
Other: ON: DE95626630; TRN: BR9532980040665
Availability:
INIS; OSTI as DE95626630
Submitting Site:
INIS
Size:
177 p.
Announcement Date:
May 15, 1995

Citation Formats

Silva, S.R. da. Production of double antibody for radioimmunoassay (sheep anti-rabbit IgG antiserum); Producao de duplo anticorpo para radioimunoensaio (antissoro de carneiro anti-IgG de coelho). Brazil: N. p., 1993. Web.
Silva, S.R. da. Production of double antibody for radioimmunoassay (sheep anti-rabbit IgG antiserum); Producao de duplo anticorpo para radioimunoensaio (antissoro de carneiro anti-IgG de coelho). Brazil.
Silva, S.R. da. 1993. "Production of double antibody for radioimmunoassay (sheep anti-rabbit IgG antiserum); Producao de duplo anticorpo para radioimunoensaio (antissoro de carneiro anti-IgG de coelho)." Brazil.
@misc{etde_38160,
title = {Production of double antibody for radioimmunoassay (sheep anti-rabbit IgG antiserum); Producao de duplo anticorpo para radioimunoensaio (antissoro de carneiro anti-IgG de coelho)}
author = {Silva, S.R. da}
abstractNote = {A second antibody (sheep anti-rabbit IgG antiserum) to be used in RIAs in which the first antibody is raised in rabbits was produced. For this production, initially the IgG was isolated from rabbit serum and purified by sodium sulphate precipitation followed by ion exchange chromatography on DEAE-cellulose. Four sheep were immunized with 500 u g of purified rabbit IgG, emulsified in Freund Complete Adjuvant and administered by multisite subcutaneous injections. These injections were repeated at 20-days intervals and blood samples (40 ml) were taken from the jugular vein 10 days after the boosts for the evaluation of the antisera title. After each four boosts a great bleeding was done by the same route. Approximately 500 ml of serum were obtained in each bleeding per animal. The antisera were evaluated by the human thyrotropin RIA developed at IPEN laboratories employing reagents provided by NIDDKD, USA. These evaluations referred to the determination of the antisera title and of the ideal concentration of carrier IgG, to the study of the kinetic of precipitation and to the confirmation of the inexistent cross-reactivity with human IgG, in comparison with a reference antiserum of know precipitation characteristics supplied by the Radioassay System Laboratories. Approximately 3,6 l of antiserum (sheep anti-rabbit IgG serum) were produced from the four sheep, which presented title and precipitation characteristics very similar to those exhibited by the imported commercial product, even presenting higher titles. The results obtained in this work indicated that it was created enough experience for the production of this biological reagent for RIA, that could be done integrally in the country in greater scale, and at a very reduced cost. (author). 81 refs, 36 figs, 33 tabs.}
place = {Brazil}
year = {1993}
month = {Dec}
}