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Proteus vulgaris

Proteus vulgaris

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Published by: api-19649313 on Nov 25, 2009
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03/18/2014

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Chapter 1
INTRODUCTION

Antibody production is important in researches, medical industry and can be indispensable tools to analyze protein functions in the broad area of life science. Antibodies are one of the most important tools for the diagnosis and components for vaccines. Animals like rabbit, mouse, guinea pig, chicken, goat, sheep, hamster and rat have been used frequently for the production of antibodies.

Antibodies are soluble proteins by B cells, which interact with antigens. Antigens are molecules capable of interacting with specific components of the immune system (Madigan, M. T. et al., 2003). Polyclonal antibodies are antibodies that are derived from many different cells or cell lines, similar to the mixture of antibodies found in sera. Polyclonal antibodies are therefore a mixture of much different specificity. This is in contrast to monoclonal antibodies which are derived from one clone (Wagner, 2007). There were researches conducted for the production and characterization of polyclonal antibodies. Polyclonal antibody production in mammals is generally associated with multiple injections of antigens, adjuvants and repeated blood sampling procedures (Hau, J. et al., 2005). Certain antibodies can be produced to protect the body from different pathogens like bacteria, fungi or viruses.

Proteus is a genus of Enterobacteriaceae, which occurs widely in humans and
animals and in the environment, and can be easily recovered from sewage, soil, garden
vegetables and many other materials.Proteus strains are often found as concomitants of
Shigella and make their appearance in the stools of patients recovering from bacillary
dysentery. They can cause many illnesses, such as kidney stones, bladder stones, peritonitis,
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septicaemia, pyelitis and infection. Also, the diagnosis of pyelonephritis and urinary tract infection can be made when thePr oteus concentration in urine is greater than 105 cells/ml. Therefore, the detection and analysis of the growth characteristics ofProteus is of importance in clinical microbiology, environmental science, food technology, toxicology and biotechnology (Tan, H. et al., 1997)

Proteus vulgaris is an opportunistic pathogen and has been associated with cases of

bacteremia, pneumonia and focal lesions. It can cause many different types of infection including urinary tract infections and wound infections and is a common cause of sinus and respiratory infections, especially in South East Asia, which can be extremely hard to eradicate in sinus and respiratory tissues (Baron, S. et al., 1996).

Proteus vulgaris is gram-negative facultative anaerobe that has a growth temperature

of 37\u00b0C. Proteus vulgaris occurs naturally in the intestines of humans and a wide variety of animal, manure, soil and polluted waters. Proteus vulgaris is highly motile and often swarm across the surface of agar plates, giving a distinct appearance different than distinct colonies normally seen.

Mice are used frequently for production of monoclonal antibodies. Their small size has generally been considered an impediment to collection of enough serum for harvesting adequate quantities of polyclonal antibodies. However, the use of mice for polyclonal antibody production may be particularly advantageous when the quantity of antigen is limited to a few or when only a limited amount of the antibody is needed. On the other hand, mouse size may no longer be a major drawback in terms of quantity of antibodies that can be harvested, as polyclonal antibodies can be harvested from ascitic fluid. A number of methods have been developed for producing ascites in mice (Cartledge et al., 1992; Kurpisz

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et al., 1988; Lacy and Voss, 1986; Mahana and Paraf, 1993; Maurer and Callahan, 1980;
Tung, 1983) with variable success.

The objectives of this study are to obtain high-titer, high-affinity antisera in a manner consistent with the welfare of the mice being immunized in order to produce polyclonal antibodies against a specific antigen which is Proteus vulgaris, to quantify the amount of antibody produced by ELISA, to characterize the antigen through molecular weight determination by Western blotting and to indicate the occurrence of a specific antigen- antibody reaction by IFAT.

Polyclonal antibodies that can be produced can be used to fight the pathogenPr oteus
vulgaris. This research can give more ideas on the interaction of the antibody and antigen

regarding Proteus vulgaris antigen. It could develop new diagnostic approach and therapeutics regardingProteus infection. The scope of the study is for the production, characterization and quantification of murine polyclonal antibodies only. It also includes characterization of highly immunogenic protein in Proteus vulgaris through Western blotting. It does not include the study of properties, application on therapeutics, study of mechanism of antibodies, further purifications and cross-reactivity of polyclonal serum generated.

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