Rapid dual-column chromatographic assay for recombinant leukocyte interferon α-2

Rybacek, Ludvik; D'Andrea, Mark; Tarnowski, S.Joseph

doi:10.1016/s0021-9673(01)85019-6

Cited by 24 publications

(8 citation statements)

References 16 publications

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“…One is that pre-column immunoextraction may eliminate RPLC solvent incompatibility since immunoassays are performed prior to LC. A second advantage is that the immunoaffinity column can concentrate antigens from a large volume of sample (Frutos and Regnier, 1993); the concentration sensitivity of the assay may therefore be improved 10-20 times (Rybacek et al, 1987). Finally, analytes that are sequestered by proteins may be captured, which is often the case in the analysis of drugs and hormones where nonspecific protein binding may interfere with determination.…”

Section: Pre-column Immunoassays Coupled To Lcmentioning

confidence: 99%

Coupling immunoassays with chromatographic separation techniques

Tang

Karnes

2000

Biomed. Chromatogr.

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Coupling immunoassays with HPLC separation techniques is becoming increasingly useful in the analysis of biological and nonbiological samples of both large and small molecules. This is because it provides both sensitivity and selectivity for molecular analysis at relatively low cost, low maintenance and with excellent potential for automation. This paper reviews application of this hyphenated approach both in the pre-column immunoextraction and post-column immunodetection modes. Systems in which immunoassays are interfaced to chromatographic separations in order to separate bound and free fractions of the immunoassay will not be included since these systems do not provide the enhanced selectivity common to hyphenated systems. Post-column immunodetection is based on various immunoassay formats such as direct detection, one-site, competitive and sandwich immunoassays. Homogeneous immunodetectors are more convenient than heterogeneous immunodectors since there are no separation and column regeneration steps involved in homogeneous immunoassays. On the other hand, heterogeneous immunoassays are generally more sensitive than homogeneous immunoassays since interfering substances are removed prior to immunodetection. Advantages and limitations for the various approaches will be discussed.

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Section: Pre-column Immunoassays Coupled To Lcmentioning

confidence: 99%

Coupling immunoassays with chromatographic separation techniques

Tang

Karnes

2000

Biomed. Chromatogr.

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“…Another type of pre-column immunoassay involves coupling an immunoaffinity column on-line with the aid of a switching valve [32][33][34][35][36][37]. The immunoaffinity columns in both cases consist of antibodies immobilized on a solid phase.…”

Section: Precolumn Immunoassays Coupled To Hplcmentioning

confidence: 99%

Combining immunoassays with chromatographic and electrophoretic separation techniques ? a review

Shahdeo

Karnes

1998

Mikrochim Acta

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Abstract. The combination of immunoassays with separation techniques such as chromatography and electrophoresis can provide both selectivity and sensitivity that is competitive with any method currently available for molecular analysis. Immunoassays can be carried out on-line and off-line, pre and post separation. The on-line post separation mode is the most promising for routine analysis because of the high throughput that can be achieved but also provides the greatest challenge with regard to compatibility of the interfaced systems. This paper reviews the various approaches that have been researched from a practical immunochemical point of view with emphasis on the special problems incurred with matrix compatibility for on-line post separation systems.

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“…A survey of literature reported by Farjam (18) showed less than thirty reports involved molecules under 1,000 molecular weight There were affinity methods for steroids such as testosterone, nortestosterone, Cortisol (19)(20)(21)(22) and carcinogenic compounds such as afflatoxins and ochratoxin (23)(24)(25)(26). Among the techniques reported, immobilized antibodies have been used for on-line sample pre-treatments in liquid and gas chromatographies (10,27), liquid chromatography/ mass spectrometry (21,29) and manual affinity chromatography for residue sample clean-up. (30).…”

Section: Columns the First Application Of Monoclonal Antibody Affinimentioning

confidence: 99%

Immunoaffinity Chromatography Applications in Pesticide Metabolism and Residue Analysis

Wong¹,

Pont²,

Johnson³

et al. 1995

ACS Symposium Series

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The imidazolinone compounds are a new class of herbicides which are environmentally safe and effective at very low application rates. Because of the shared imidazolinone ring structure amongst several herbicides, generic antibodies were prepared which recognized all the compounds in this class (1). These antibodies are useful not only for developing immunoassays for the detection of low levels of residues in soil or in plant extracts, they are also useful for preparing antibody affinity columns used for sample clean-up in metabolism and residue studies. Imidazolinone-compound containing extracts from wheat plant, corn grain, corn fodder, goat urine, and goat kidney tissue were tested on the antibody columns. It was found that the analytes were bound to the antibody column after some simple sample preparation procedures. The bound analytes were easily eluted with a solution of 30% methanol in water. It was further demonstrated that metabolites of imidazolinone compounds which retain the imidazolinone ring structure can be purified through a simple antibody affinity chromatography procedure and be identified thereafter by mass spectrometry. A comparison of mono-and poly-clonal antibody columns indicates that the monoclonal antibody with its uniform affinity and more restricted binding epitope is better for antibody affinity chromatographies. A lower amount of matrix is bound to the monoclonal antibody column, thus producing a cleaner sample than a polyclonal antibody column.The specificity of antibodies has been utilized extensively in immunoassays where analytes can be quantified in relatively crude matrices. This property can be further harnessed in affinity chromatography. Preparation of monoclonal antibodies allows an unlimited supply of mono-specific antibodies for large scale production of affinity 0097-6156/95/0586-0235$12.00/0

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Rapid dual-column chromatographic assay for recombinant leukocyte interferon α-2

Cited by 24 publications

References 16 publications

Coupling immunoassays with chromatographic separation techniques

Coupling immunoassays with chromatographic separation techniques

Combining immunoassays with chromatographic and electrophoretic separation techniques ? a review

Immunoaffinity Chromatography Applications in Pesticide Metabolism and Residue Analysis

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