Immunoaffinity chemistry

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. 

Hodgson, R. J., Brook, M. A., and Brennan, J. D., CapiUary-scale monolithic immunoaffinity columns for immunoextraction with in-line laser-induced fluorescence detection. Analytical Chemistry 77(14), 4404-4412, 2005. 

Because of the time and effort needed to develop immunoaffinity methodology (4 to 8 weeks typically needed to generate a purified antibody), immunoaffinity separations are usually reserved for the more difficult separation problems in analytical chemistry. The shortened time scale of drug discovery precludes their routine day-to-day use. The power of immunosepara-tions in combination with mass spectrometry could see wider use as clever solutions to difficult problems are needed. 

Figure 4 Example of an approach to immunoaffinity recognition in CZE using an FL probe Fab fragment of the Ab (61). CIEF analysis of crude TR-Fab and its complexes with met-rhGH. APCE was done with Pharmalyte 3-10 and using TR-Fab before purification as an affinity probe with (a) TTA-BSA buffer and (b) met-rhGFl (1 p,g/ml) as samples. The free TR-Fab and the complexes are marked by and, respectively. (Reproduced with permission from the copyright holder, American Chemical Society and Analytical Chemistry.)...

Tao, W.-J. M. A. Quan, Gritsenko, D. G. Camp, M. E. Monreo, R. J. Moore, and R. D. Smith. 2005. Human plasmaN-Glycoprotein analysis by immunoaffinity substraction, hydrazine chemistry, and mass spectrometry. J. Proteom. Res. 4, 2070-2080. 

Cai, J., and Henion, J. (1996) On line immunoaffinity extraction coupled column capillary liquid chromatography/tandem mass spectrometry trace analysis of LSD analogs and metabolites in human urine. Analytical Chemistry, 68, 72 78. 

Driedger, D.R., and Spoms, P. (2001) Immunoaffinity sample purification and MALDI TOF MS analysis of a solanine and a chaconine in serum. Journal of Agricultural and Food Chemistry, 49, 543 548. 

Nakade, S., Rhee, S.K., Hamanaka, H., and Mikoshiba, K. (1994) Cyclic AMP-dependent phosphorylation of an immunoaffinity-purified homotetrameric inositol 1,4,5-trisphosphate receptor (type I) increases Ca flux in reconstituted lipid vesicles. Journal of Biological Chemistry, 269 6735-6742. 

Nilsson, S., Lager, C., LaureU, T., and Bimbaum, S., Thin-layer immunoaffinity chromatography with bar code quantitation of C-reactive protein. Analytical Chemistry, 67, 3051-3056, 1995. 

In the frame of mycotoxin analysis, it is worth remembering that immunoaffinity columns are used for enrichment of samples to be analyzed by solution chemistry or by liquid chromatography. The use of immunoaffinity in this case allows a one-step removal of possible interfering agents from the sample. 

Hage, D.S. Clarke, W. Immunoaffinity chromatography in clinical analysis. In Separation Techniques in Clinical Chemistry Aboul-Enein, H.Y., Ed. Marcel Dekker New York, 2003 361-387. 

Fenaille, E, Tabet, J.C., Guy, PA. (2002) Immunoaffinity purification and characterization of 4-hydroxy-2-nonenal-and malondialdehyde-modified peptides by electrospray ionization tandem mass spectrometry. Analytical Chemistry, 74, 6298-6304. 

Becher, E, et ai. (2007) Detection of functional ricin by immunoaffinity and liquid chromatography-tandem mass spectrometry. Analytical Chemistry, 79, 659-665. 

Immunoaffinity extraction is a technique often adopted in clinical chemistry. Immu-noaffinity supports contained polyclonal antibodies covalently immobilized on sephar-ose, agarose, or silica supports. Two different kinds of supports, crushed sol-gel monoliths and sol-gel-coated highly porous silica particles, were applied to the extraction of 16 sulfonylureas in food and natural water followed by high-performance liquid chromatography-ultraviolet/diode-array detection (LC-UV/DAD) [32]. 

Thevis, M., Thomas, A., Delahaut, P. (2005) Qualitative determination of synthetic analogues of insuhn in human plasma by immunoaffinity purification and liquid chromatography-tandem mass spectrometry for doping control purposes. Analytical Chemistry, Tl, 3579-3585. 

---