Immunoaffinity

Another example of vims clearance is for IgM human antibodies derived from human B lymphocyte cell lines where the steps are precipitation, size exclusion using nucleases, and anion-exchange chromatography (24). A second sequence consists of cation-exchange, hydroxylapatite, and immunoaffinity chromatographies. Each three-step sequence utilizes steps based on different properties. The first sequence employs solubiUty, size, and anion selectivity the second sequence is based on cation selectivity, adsorption, and selective recognition based on an anti-u chain IgG (24). 

Immunoaffinity chromatography utilizes the high specificity of antigen—antibody interactions to achieve a separation. The procedure typically involves the binding, to a soHd phase, of a mouse monoclonal antibody which reacts either directly with the protein to be purified or with a closely associated protein which itself binds the product protein. The former approach has been appHed in the preparation of Factor VIII (43) and Factor IX (61) concentrates. The latter method has been used in the preparation of Factor VIII (42) by immobilization of a monoclonal antibody to von WiHebrand factor [109319-16-6] (62), a protein to which Factor VIII binds noncovalenfly. Further purification is necessary downstream of the immunoaffinity step to remove... 

For detection residue amounts of tetracyclines in dairy products widely used methods FIPLC, immunoaffinity chromatography, kinetic spectrophotometry, which are expensive and complicated. 

T.M. Phillips and B.F. Dickens, Affinity and Immunoaffinity Purification Techniques, Eaton Publishing, Natick MA, 2000. ISBN 1881299228. 

W. Haasnoot, M. E. Ploum, R. J. A. Paulussen, R. Schilt and F. A. Huf, Rapid determination of clenbuterol residues in urine by high perfoi mance liquid cliromatography with on-line automated sample processing using immunoaffinity chromatography , 7. Chromatogr. 519 323-335 (1990). 

A. Faijam, A. E. Brugman, A. Soldaat, P. Timmerman, H. Fingerman, G. J. de Jong, R. W. Frei and U. A. Th Brinkman, Immunoaffinity precolumn foi selective sample preti eatment in column liquid cliromatography immunoselective desoiption , Chromatographia 31 469-477 (1991). 

G. S. Rule, A. V. Moi dehai and J. Henion, Determination of carbofuran by on-line immunoaffinity cliromatography with coupled-column liquid chromatography/mass spectrometiy . Aim/. Chem. 66 230-235 (1994). 

J. Cai and J. Henion, Quantitative multi-residue determination of /3-agonists in bovine urine using on-line immunoaffinity extr action-coupled column packed capillary liquid cliromatogr aphy-tandem mass spectr ometry , 7. Chromatogr. 691 357-370 (1997). 

C. S. Creaser, S. J. Feely, E. Houghton and M. Seymour, Immunoaffinity cliromatogr-a-phy combined on-line with liigh performance liquid cliromatogr aphy-mass spectr ome-tr y for the determination of corticosteroids , 7. Chromatogr. 794 37-43 (1998). 

Immunoaffinity extraction combined on-line with LC in conjunction with MS (108 -110) or tandem MS (111, 112) has also been demonstrated for the determination of analytes in biological fluids. Obviously, such systems offer a very high... 

Figure 11.13 (a-c) Immunoaffinity exti action-SPE-GC-FID ti aces of (a) HPLC-grade water (b) urine (c) urine spiked with /3-19-noitestosti one (0.5 p.g/1) or norethindrone and norgestrel (both 4 p.g/1) (d) SPE-GC-FID ti ace of urine. Reprinted from Analytical Chemistry, 63, A. Faijam et al., Direct inti oduction of large-volume urine samples into an on-line immunoaffinity sample pretreatment-capillary gas cliromatography system, pp. 2481-2487,1991, with permission from the American Chemical Society. 

Analogous to two-dimensional LC, the on-line coupling of LC and CE has been carried out both in the heart-cut and the comprehensive mode. In a heart-cut LC-CE study, a protein G immunoaffinity LC column was used to selectively preconcentrate insulin from serum (171). A 1 p.1 elution plug comprising the insulin was switched on-line to the CE system where a part was injected into the capillary for final separation. With CE, efficient separations can be obtained in a... 

B. Johansson, Simplified quantitative determination of plasma phenytoin on-line pre-column high-perfor mance liquid immunoaffinity cliromatogr aphy with sample pre-purification , J. Chromatogr. 381 107-113 (1986). 

W. Haasnoot, R. Scliilt, A. R. M. Hamers, F. A. Huf, A. Farjam, R. W. Frei and U. A. Th Brinkman, Determination of /3-19-nortestosterone and its metabolite a- 9-nortestosterone in biological samples at the sub parts per billion level by high-performance liquid cliromatography with on-line immunoaffinity sample pretreatment , J. Chromatogr. 489 157-171 (1989). 

A. Farjam, N. C. van de Merbel, A. A. Nieman, H. Lingeman and U. A. Th Brinkman, Determination of aflatoxin Ml using a dialysis-based immunoaffinity sample pretreat-ment system coupled on-line to liquid cliromatography , ]. Chromatogr. 589 141-149(1992). 

V. Kircher and H. Parlar, Determination of teti ahydrocannabinol from human saliva by tandem immunoaffinity cliromatography-liigh-peiformance liquid cliromatogra-phy , ]. Chromatogr. B 677 245-255 (1996). 

E. Davoli, R. Fanelli and R. Bagnati, Purification and analysis of dmg residues in urine samples by on-line immunoaffinity cliromatography/bigh-performance liquid cliro-matography/continuos-flow fast atom bombardment mass spectrometry . Anal. Chem. 65 2679-2685 (1993). 

G. S. Rule and J. D. Henion, Determination of drugs from urine by on-line immunoaffinity cliromatography-high-performance liquid cliromatography-mass specU ometiy , J. Chromatogr. 582 103-112 (1992). 

J. Cai and J. Henion, On-line immunoaffinity exti action-coupled column capillary liquid cliromatography/tandem mass specb omeb y trace analysis of LSD analogs and metabolites in human urine . Anal. Chem. 68 72-78 (1996). 

A. Earjam, J. J. Vreuls, W. J. G. M. Cuppen, U. A. Th Brinkman and G. J. de Jong, Duect introduction of large-volume urine samples into an on-line immunoaffinity sample pre-treatment-capillary gas chr omatogr aphy system . Awn/. Chem. 63 2481-2487 (1991). 

E. J. Cole and R. T. Kennedy, Seleaive preconcenti ation for capillary zone electrophoresis using protein G immunoaffinity capillary chi omatography . Electrophoresis 16 549-556(1995). 

The low selectivity of the SPE columns currently in use can be increased with more selective sorbents such as the immunosorbents, which have been quite extensively used in SPE-LC (72). Immunoaffinity-based solid-phase extraction (lASPE) sorbents have also been used in coupled gas chromatography for determining... 

Figure 13.20 GC-FID chromatograms of an exuact obtained by (a) SPE and, (b) lASPE of 10 ml of municipal waste water, spiked with 1 p.g 1 of seven s-triazines (c) represents a blank mn from lASPE-GC-NPD of 10 ml of EIPLC water. Peak identification is as follows 1, ati azine 2, terbuthylazine 3, sebuthylazine 4, simetiyn 5, prometiyn 6, terbutiyn 7, dipropetiyn. Reprinted from Journal of Chromatography, A 830, J. Dalliige et al, On-line coupling of immunoaffinity-based solid-phase exUaction and gas chi-omatography for the determination of 5-triazines in aqueous samples , pp. 377-386, copyright 1999, with permission from Elsevier Science.

J. Dalliige, T. Hankemeier, J. J. Vreuls and U. A. Th Brinkman, On-line coupling of immunoaffinity-based solid-phase extr action and gas chr omatography for the determination of 5-triazines in aqueous samples , 7. Chromatogr. 830 377-386 (1999). 

Human interleukin 2, a 133-residue protein, has been separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. Most of the heterogeneity has been attributed to variations in glycosylation of the threonine residue in position 3 of the polypep-... 

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