Immunological-Chromatographic Analysis

Janis, Linda J.; Regnier, Fred E.

doi:10.1016/s0021-9673(01)94003-8

Cited by 48 publications

(18 citation statements)

References 17 publications

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“…The fact that antibodies target a single structural feature (an epitope) of an antigen is a major strength [5]. Immunoaffinity chromatography (IAC) [6] and immunological assays have a level of structural selectivity not seen in other analytical and preparative systems. It is frequently the case that an immunosorbent will purify an antigen from a complex biological sample several thousand fold in a single step [7].…”

Section: Introductionmentioning

confidence: 99%

Screening antibody and immunosorbent selectivity by two‐dimensional liquid chromatography‐MS/MS (2‐D LC‐MS/MS)

Cho

Jung

Regnier

2010

J of Separation Science

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Selectivity of both peptide-and glycan-targeting antibodies was examined by 2-D LC-MS/ MS. Proteins selected from biological extracts immunospecifically in a first chromatography dimension using antibodies immobilized by either covalent coupling or adsorption to protein G were desorbed with a denaturing mobile phase and transferred to a 1.5 mm nonporous particle RP chromatography (NP-RPC) column in a second dimension. Protein peak capacity of the NP-RPC column was approximately 50. Peaks collected from the RPC column were tryptic digested and the peptide fragments were identified by MALDI-MS/ MS. The objective of this analytical strategy was to discriminate between protein antigens and nonantigens through identification of their peptides, leading to an evaluation of the selectivity of antibodies and immunosorbents. Quantification of the relative amount of antigen and nonantigen species captured by immunosorbents was achieved by absorbance, along with the likely capture mechanism. A limitation of the approach was in discriminating between isoforms of an antigen in which neither the antibody nor the LC-MS system targeted the differentiating feature in the isoforms.

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Section: Introductionmentioning

confidence: 99%

Screening antibody and immunosorbent selectivity by two‐dimensional liquid chromatography‐MS/MS (2‐D LC‐MS/MS)

Cho

Jung

Regnier

2010

J of Separation Science

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“…Simultaneous determination of flunitrazepam and its metabolites in human urine by UV absorbance was published by Deinl et al, (1997). Some assays involving protein analytes have utilized immunoassay coupled with size exclusion (Riggin et al, 1993) or ion-exchange chromatography (Janis and Regnier, 1988;Janis et al, 1989).…”

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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“…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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Immunological-Chromatographic Analysis

Cited by 48 publications

References 17 publications

Screening antibody and immunosorbent selectivity by two‐dimensional liquid chromatography‐MS/MS (2‐D LC‐MS/MS)

Screening antibody and immunosorbent selectivity by two‐dimensional liquid chromatography‐MS/MS (2‐D LC‐MS/MS)

Coupling immunoassays with chromatographic separation techniques

Combining immunoassays with chromatographic and electrophoretic separation techniques ? a review

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