Immunoaffinity chromatograph

A rapid tic immunoaffinity chromatographic method has been reported for quantitation in serum of an acute phase reactant, C-reactive protein (CRP), which can differentiate between viral and bacterial infections. 

In the production process of Kogenate , BHK cells are cultivated in perfusion bioreactors up to 500 L in size. The first step of purification involves anion exchange chromatography. After that, an immunoaffinity chromatographic step and gel filtration are carried out, followed by another immunoaffinity and another anion exchange chromatography step (Boedeker, 1992 Bhattacharyya et al., 2003). 

Lawrence, J.F. Menard, C. Determination of clenhuterol in heef-liver and muscle-tissue using immunoaffinity chromatographic cleanup and liquid-chromatography with ultraviolet absorbency detection. J. Chromatogr. 1997, 696, 291-297. 

The first row includes assorted methods where the analyte binds to the antibody, is physically separated from the sample matrix, and then measured. These include immunoaffinity solid-phase extraction, where after separation of the bound antigen from the sample matrix, the antigen is subsequently released and measured, and also several methods that depend on two antibodies binding independently to the analyte, such as the immunoaffinity chromatographic methods behind several home-test diagnostic kits such as pregnancy tests. 

Sharman, M. and Gilbert, J., Automated aflatoxin analysis of foods and animal feeds using immunoaffinity column clean-up and high-performance liquid chromatographic determination, /. Chromatogr., 543, 220, 1991. 

Wellferon was the tradename given to one of the first such approved products. Produced by large-scale mammalian (lymphoblastoid) cell cultures, the crude preparation undergoes extensive chromatographic purification, including two immunoaffinity steps. The final product contains nine IFN-a subtypes. 

Immunoaffinity procedures can be performed either on-line or off-line, and can be coupled to chromatographic systems [ 118,119] or even to immunoassays [120]. Many examples can be found in the literature regarding the use of immunoaffinity extraction of drugs and pharmaceuticals from biological matrices, as well as of organic pollutants such as pesticides from environmental samples [115,121-124]. 

Both small and large analytes can be determined using direct detection in lAC. Additionally it is possible to use this technique either separately or in combination with other chromatographic techniques [1]. If this technique is performed as part of HPLC system the method can be referred as high performance immunoaffinity chromatography. 

A specific cleanup procedure based on immunoaffinity chromatography with polyclonal antibodies has been described by Gude et al. (50) for the gas chromatographic determination of chloramphenicol in swine tissues. In this method, tissue sample is extracted with acetonitrile/4% sodium chloride (1 1) Following centrifugation, the supernatant is purified with n-hexane, and chloram-... 

During recent years, however, the distinction between these two types of methods has become less clear because of the improved methods of sample cleanup that allow selective isolation of groups of compounds. Mixed-mechanism solid-phase extraction procedures and multi-immunoaffinity techniques are clear examples of liquid chromatographic developments that have contributed greatly to the current state of the art within residue analysis. 

A Patey, M Sharman, J Gilbert. Liquid chromatographic determination of aflatoxins level in peanut butter using an immunoaffinity columns clean-up. method international collaborative trials. J Ass Off Anal Chem 74(1) 76-81, 1991. 

M Sharman, S Mac Donald, J Gilbert. Automated liquid chromatographic determination of ochratoxin A in cereals and animal products using immunoaffinity column clean-up. J Chiom 603 285-289, 1992. 

A Pittet, D Tornare, A Huggett, R Viani. Liquid chromatographic determination of ochratoxin A in pure and adulterated soluble coffee using an immunoaffinity column cleanup procedure. J Agric Food Chem 44 3564-3569,1996. 

Lawrence, J.F., C. Menard, M.-C. Hennion, et al. 1996. Use of immunoaffinity chromatography as a simplified cleanup technique for the liquid chromatographic determination of phenylurea herbicides in plant material. J. Chromatogr. A 732 277-281. 

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