ISSN 1214-0287 (on-line)
ISSN 1214-021X (printed)

Volume 7 (2009), No 3, p 115-121

Monoclonal and polyclonal antibodies production - preparation of potent biorecognition element

Miroslav Pohanka

Address: Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defense, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic

Received 14th June 2009.
Revised 3rd August 2009.
Published online 3rd September 2009.

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Antibodies are macromolecular structures with a high affinity to a molecule called the antigen. Though the natural aim of antibodies is the protection of the body from pathogens, they have been found to play a useful part in a number of specific treatments and diagnoses. This wider role for antibodies extends to assays where antibodies provide a recognition capability improving available physical methods. Photometrical methods such as ELISA or flow cytometry and electrochemical, optical or piezoelectric immunosensors (biosensors) are typical examples. The present review summarizes facts about the parameters and the production of antibodies. The structure of immunoglobulins, and the production and purification of monoclonal and polyclonal antibodies are described in four chapters. The review was written in order to collect the available knowledge on antibodies into one study improving orientation in this field for anyone wanting to construct immunoassays.

antibody; immunoglobulin; biosensor; polyclonal; monoclonal; immunoassay

Amrouche T, Boutin Y, Moroni O, Kheadr E, Fliss I: Production and characterization of anti-bifidobacteria monoclonal antibodies and their application in the development of an immuno-culture detection method. J Microbiol Methods 65:159-170, 2006.

Anderson GP, King KD, Faffney KL, Johnson LH: Multi-analyte interrogation using the fiber optic biosensor. Biosens Bioelectron 14:771-777, 2000.

Akerstrom B, Nielsen E, Bjorck L: Definition of the IgG- and the albumin-binding regions of streptococcal protein G. J Biol Chem 262:13388-13391, 1987.

Bergmann-Leitner ES, Mease RM, Duncan EH, Khan F, Waitumbi J, Angov E: Evaluation of immunoglobulin purification methods and their impact on quality and yield of antigen-specific antibodies. Malar J 7:129, 2008.

Crowley EL, O'Sullivan CK, Guilbault GG: Increasing the sensitivity of Listeria monocytogenes assays: evaluation using ELISA and amperometric detection. Analyst 124:295-299, 1999.

Dwek RA, Lellouch AC, Wormald MR: Glycobiology: "the function of sugar in the IgG molecule". J Anat 187:279-292, 1995.

Emanuel PA, Dang J, Gebhardt JS, Aldrich J, Garber EAE, Henrieta K, Stopa P, Valdes JJ, Schultz AD: Recombinant antibodies: a new reagent for biological agent detection. Biosens Bioelectron 14:761-770, 2000.

Erhard MH, Kuhlmann R, Szinicz L, Losch U: Detection of the organophosphorus nerve agent soman by an ELISA using monoclonal antibodies. Arch Toxicol 64:580-585, 1990.

Externest D, Mackelein B, Schmidt MA, Frey A: Correlations between antibody immune responses at different mucosal effector sites are controlled by antigen type and dosage. Infect. Immun. 68:3830-3839, 2000.

Goryacheva IY, DeSaeger S, Delmulle B, Lobeau M, Eremin SA, Barna-Vetro I, Van Peteghem C: Simultaneous non-instrumental detection of aflatoxin A using a clean-up tandem immunoassay column. Anal Chim Acta 590:118-124, 2007.

Heldman E, Balan A, Horowitz O, Ben-Zion S, Torten M: A novel immunoassay with direct relevance to protection against organophosphate poisoning. FEBS Lett 180:243-248, 1985.

Hemachandra S, Kamboj K, Copfer J, Pier G, Green LL, Schreiber JR: Human monoclonal antibodies against Pseudomonas aeruginosa lipopolysaccharide derived from transgenic mice containing megabase human immunoglobulin loci are opsonic and protective against fatal pseudomonas sepsis. Infect Immun 69:2223-2229, 2001.

Kohler G, Milstein C: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495-497, 1975.

Kolosova AY, Shim WB, Yang ZY, Eremin SA, Chung DH: Direct competitive ELISA based on a monoclonal antibody for detection of aflatoxin B1. Stabilization of ELISA kit components and application to grain samples. Anal Bioanal Chem 384:286-294, 2006.

Kowal-Bielecka O, Kowal K, Distler O, Rojewska J, Bodzenta-Lukaszyk A, Michel BA, Gay RE, Gay S, Sierakowski S: Cyclooxygenase-and lipoxygenase-derived eicosanoids in bronchoalveolar lavage fluid from patients with scleroderma lung disease: an imbalance between proinflammatory and antiinflammatory lipid mediators. Arthritis Rheum 52:3783-3791, 2005.

Laegreid W, Hoffman M, Keen J, Elder R, Kwang J: Development of a blocking Enzyme-Linked Immunosorbent Assay for detection of serum antibodies to O157 antigen of Escherichia coli. Clin Diagn Lab Immunol 5:242-246, 1998.

Little SF, Leppla SH, Friedlander AM: Production and characterization of monoclonal antibodies against the lethal factor component of Bacillus anthracis lethal toxin. Infect Immun 58:1606-1613, 1990.

Martinez JM, Martinez MI, Suarez AM, Hezzanz C, Casaus P, Cintas LM, Rodriguez JM, Hernandez PE: Generation of polyclonal antibodies of predetermined specificity against pediocin PA-1. Appl Environ Microbiol 64:4536-4545, 1998.

Mendez MJ, Green LL, Corvalan JR, Jia XC, Maynard-Currie CE, Yang XD, Gallo ML, Louie DM, Lee DV, Erikson KL, Luna J, Roy CM et al.: Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response in mice. Nat Genet 15:146-156, 1997.

Morris TJ, Stanley EF: A simple method for immunocytochemical staining with multiple rabbit polyclonal antibodies. J Neurosci Methods 127:149-155, 2003.

Nimmo GR, Lew AM, Stanley CM, Steward MW: Influence of antibody affinity on the performance of different antibody assays. J Immunol Methods 72:177-187, 1984.

Parr DM: Fragments produced by digestion of human immunoglobulin G subclasses with pepsin in urea. Biochem J 165:303-308, 1977.

Pasupathi P, Bakthavathsalam G, Saravanan G, Devaraj A: Changes in CD4+ cell count, lipid profile and liver enymes in HIV infection and AIDS patients. J Appl Biomed 6:139-145, 2008.

Pohanka M: Evaluation of immunoglobulin production during tularaemia infection in BALB/c mouse model. Acta Vet Brno 76:579-584, 2007.

Pohanka M, Skladal P: Piezoelectric immunosensor for the direct and rapid detection of Francisella tularensis. Folia Microbiol 52:325-330, 2007.

Pohanka M, Skladal P: Electrochemical biosensors - principles and applications. J Appl Biomed 6:57-64, 2008.

Pohanka M, Pavlis O, Skladal P: Diagnosis of tularemia using piezoelectric biosensor technology. Talanta 71:981-985, 2007.

Pohanka M, Pavlis O, Kroca M: ELISA detection of Francisella tularensis using polyclonal and monoclonal antibodies. Def Sci J 58:698-702, 2008.

Pohanka M, Malir F, Roubal T, Kuca K: Detection of aflatoxins in Capsicum spice using an electrochemical immunosensor. Anal Lett 41:2344-2353, 2008b.

Somodevilla-Torres MJ, Hillyard NC, Morton H, Alewood D, Halliday JA, Alewood PF, Vesey DA, Walsh MD, Cavanagh AC: Preparation and characterization of polyclonal antibodies against human chaperonin 10. Cell Stress Chaperones 5:14-20, 2000.

Tejral G, Panyala NR, Havel J: Carbon nanotubes: toxicological impact on human health and environment. J Appl Biomed 7:1-13, 2009.

Ventury M, Rimon A, Gerchman Y, Hunte C, Padan E, Michel H: The monoclonal antibody 1F6 identifies a pH-dependent conformational change in the hydrophilic NH2 terminus of NhaA Na+/H+ antiporter of Escherichia coli. J Biol Chem 18:4734-4742, 2000.

Ayyar BV, Arora S, Murphy C, O'Kennedy R. Affinity chromatography as a tool for antibody purification. Methods. 56: 116-129, 2012.

Puertas S, Batalla P, Moros M, Polo E, Del Pino P, Guisan JM, Grazu V, De La Fuente JM. Taking advantage of unspecific interactions to produce highly active magnetic nanoparticle antibody conjugates. ACS Nano. 5: 4521-4528, 2011.

Li QF, Tang DP, Tang JA, Su BL, Huang JX, Chen GN. Carbon nanotube-based symbiotic coaxial nanocables with nanosilica and nanogold particles as labels for electrochemical immunoassay of carcinoembryonic antigen in biological fluids. Talanta. 84: 538-546, 2011.

Pohanka M, Vlcek V, Kuca K, Bandouchova H, Pikula J. Pesticide sorption in typical Central European soils evaluated using a photometric microplate assay based on acetylcholinesterase inhibition. J Appl Biomed. 8: 41-46, 2010.