Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G
Authors
Abstract:
Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme and F(ab')2 fragment was purified by gel filtration separation method. For production of polyclonal antibody, rabbit was immunized by purified F(ab')2 and antibody production was investigated by enzyme-linked immunosorbent assay. Purified anti-IgG F(ab')2 was conjugated with fluorescein isothiocyanate. Ion exchange chromatography purification yielded 38 mg of human IgG antibody. The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 25-30 kDa molecular weight (MW) and 50-kDa respectively and a distinct band with 150 kDa MW. The results of non-reduced SDS-PAGE for determining the purity of F(ab')2 fragment showed one band in 90 kDa and a band in 150 kDa MW position. Purification by Ion exchange chromatography method resulted about 12 mg rabbit polyclonal antibody. Flow cytometry showed generated polyclonal antibody had an acceptable activity compared to commercial antibody. Taking together, purified IgG F(ab')2 and polyclonal anti-IgG F(ab')2 are useful tools in biomedical and biochemical researches and diagnostic kits.
similar resources
Stimulation of Camel Polyclonal Antibody against Human T cell Immunoglobulin and Mucin 3
Background: T cell Immunoglobulin, Mucin (TIM)-3, is a type I transmembrane glycoprotein belonging to TIM family. This receptor expresses on T helper type 1 (Th1) cells that binds to galectin-9 (Gal9); inducing an inhibitory signal. As a result, apoptosis of Th1 cells occurs and cytotoxicity of CD8 T cells becomes evident in vitro. Therefore, this immunomodulatory molecule may ...
full textA methodological approach for production and purification of polyclonal antibody against dog IgG
Antibodies are a class of biomolecules that has an important role in the immune system and lots of applications in biotechnological methods and in pharmaceutics. Production and purification of antibodies in laboratory animals is one of the first ways to manufacture of these prominent tools. The obtained antibodies from these process could be used in various types of bioassay techniques such as ...
full textPolyclonal Antibody against Recombinant Nucleoprotein of the Influenza A Virus (H1N1); Production and Purification
Background and Aims: Influenza is an acute respiratory illness that is caused by a virus belonging to Orthomyxoviridae family. This virus spreads rapidly every year in cold season and leads to morbidities and mortalities especially in adults and children, which causes billions of dollars of economic losses. Accordingly, development of a rapid, sensitive and inexpensive laboratory diagnosis base...
full textproduction and purification of polyclonal antibody against melatonin hormone
nowadays immunochemical techniques have played a very important and valuable role in quantitative and qualitative assays of liquid compounds of the body. producing antibody against immunogenes is the first step to make immunochemical kits. in this study production and purification of polyclonal antibody against melatonin has been considered. this hormone which has several important functions in...
full textproduction of monoclonal antibody against human immunoglobulin
immunoglobulin e is one of the five classes of immonoglobulins that plays an important role in allergic diseases. production of monoclonal antibodies by a single clonotype against different epitopes of immunoglobulin e has high priority in development of diagnostic kits. in this study, an attempt was made to produce monoclonal antibodies against human immunoglobulin e. balb/c mice were immunize...
full textMy Resources
Journal title
volume 8 issue 4
pages 307- 312
publication date 2017-12-15
By following a journal you will be notified via email when a new issue of this journal is published.
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023