Production of Polyclonal Antibodies

An alternative method is to inject the gel slice that contains the protein of choice into the rabbit. This is particularly useful in 

Substances with MW 5 kd are rarely antigen. To trigger the formation of antibodies, peptides or even smaller molecules thus must be coupled to a carrier or linked into high-molecular adducts. BSA or the glycoprotein KLH (keyhole limpet hemocyanin) serve as carriers. 

Parker, X, et al. (1986). New Hydrophilicity Scale Derived from HPLC Peptide Retention Data Correlation of Predicted Surface Residues with Antigenicity and X-ray Derived Accessible Sites, Biochemistry 25 5425-5432. 

Posnett, D., et al. (1988). A Novel Method for Producing Anti-peptide Antibodies, J. Biol. Chem. 263 1719 1725. 

Seguola, R, et al. (1984). Antibodies Against y-aminobutyric Acid Specificity Studies and Immuno-cytochemical Results, Proc. Natl. Acad. Sci. USA 81 3888 3892. 

X (1988). Antibodies for Phosphotyrosine Analytical and Preparative Tool for Tyrosyl-phosphorylated Proteins, Anal. Biochem. 172 1—7. 

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Gobbo SP, Meirelles, CF (2003) Production of polyclonal antibody for ciprofloxacin detection. Proceedings of the British Society of Animal Science, York... 

Figure 4.3 Production of polyclonal antibodies from horse antisera. Source Walsh G. Biopharmaceuticals Biochemistry and Biotechnology, Wiley, Hoboken NJ, 1998.)...

The production of polyclonal antibodies by the immunization of animals is a method that has been used for more than a century (1). Hybridoma technology was the next development, allowing the production of monoclonal antibodies (2). However, hybridoma technology has some limitations, some resulting from the instability of the aneuploid cell lines, but most of all, difficulties in producing human antibodies especially to toxic or highly conserved antigens (3). 

Several qualitative and quantitative immunochemical methods and their application to the analysis of environmental samples have been described for OP insecticides, a family that includes widely used pesticides such as azinphos-ethyl/methyl, dichlorvos, fenitrothion or fenthion, malathion, mevinphos, and parathion. Mercader and Montoya202 produced monoclonal antibodies against azinphos-methyl and developed an ELISA that was used for the analysis of water samples from different sources, reaching detectability levels near 0.05 pg I. Watanabe et al.203 reported the production of polyclonal antibodies and ELISA procedures to analyze fenitrothion in river, tap, and mineral water (LOD = 0.3 pg L ). Banks et al.204 produced polyclonal antibodies against dichlorvos, an organophosphate insecticide used for stored grain, which also cross-reacts with fenitrothion. Nishi et al.205 reported the first immunoassay for malathion. Residues of this insecticide have... 

These immunoassays rely on the production of polyclonal antibodies to host cell and medium proteins which represent the most probable final... 

The production of polyclonal antibodies usually requires a relatively large amount of a pure protein, whereas the production of monoclonal antibodies may require smaller amounts of an impure protein. Explain this in terms of the method of production of these two types of antibodies. 

This chapter deals with the application of immunological techniques to biochemical and molecular biological problems. We discuss the production of polyclonal antibodies and then a range of techniques that depend on the high specificity of antibody-antigen interactions. 

Fig. 2.22. Antibody response to an antigen and the production of polyclonal antibodies. Among the four B cells shown, antibodies produced by two bind different regions of the antigen, and are stimulated to multiply.

Kurihara, N., Yanagisawa, H., Jin, Z., and Wada, O., Production of polyclonal antibodies against l,l -ethyliden bis[L-tryptophan] (EBT), a potential contaminant causing eosinophilia-myalgia syndrome (EMS), Toxicol. Lett., 66,231,1993. 

Figure 1 Schematic of production of biological antibodies, (a)-(c) illustrate production of polyclonal antibody, (a) and (d)-(h) illustrate production of monoclonal antibody, (a) inoculation (b) collect blood (c) extract antibodies (d) extract spleen cells (e) cells fused to form hybridomas (f) best antibody-producing hybridoma selected (g) hybridoma cloned (h) antibody produced and collected in cell culture.

For the generation of polyclonal antibodies, the immunogen is applied to a vertebrate (rabbit, sheep, etc.), usually with the aid of adjuvants which enhance immunogenicity, by a mechanism that is not fully understood. The production of polyclonal antibodies is not very reproducible. For this reason, antibody concentration (titer). 

There is a clear need for a specific labelling method for carbohydrates on heterogenous fibres. Antibodies can be used as specific markers for carbohydrates. Oligosaccharides derived from xylan by acid hydrolysis (10) and from xyloglucan by cellulase hydrolysis (//) have been used as antigens for production of polyclonal antibodies. Polymeric carbohydrates have also been used as antigens for antibody production (12-14). 

Latchinian-Sadek L, Ibrahim RK (1991) Flavonol ring B-specific 0-glucosyltransferases purification, production of polyclonal antibodies, and immunolocalization. Arch Biochem Biophys 289 230-236... 

From the serum of an animal, only polyclonal antibodies can be obtained, i.e. a mixture of different antibody species in different concentrations. Drawbacks of polyclonal antibodies are their critical long-term supply and a lack of reproducibility from batch to batch. The composition of a serum changes all the time and the sera of two individuals are completely different in terms of antibody concentration and their selectivity. Hence, polyclonal antibodies cannot be considered as defined reagents. On the other hand, the production of polyclonal antibodies is much cheaper than any other method and hence is often the economically most sensible way to obtain an immunoreagent. 

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