Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling.

Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Research Abstract Details 

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Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Abstract Text:

Kemmons A Tubbs,Urban A Kiernan,Eric E Niederkofler,Dobrin Nedelkov,Allan L Bieber,Randall W Nelson,

This report addresses the need for additional assays for human resistin (hRES) by developing a rational progression of the mass spectrometric immunoassay to incorporate recombinant proteins. The recombinant-based hRES mass spectrometric immunoassay (RES-MSIA) was initially developed for the qualitative analysis of the human resistin homodimer from normal (healthy) plasma samples. The method involved selective extraction and detection of both endogenous and recombinant resistant proteins. RES-MSIA was then applied to the rigorous quantification of resistin. The resistin standard addition curve was constructed from serially diluted concentrations of rhRES using endogenous hRES, inherent in the human plasma, as the internal reference standard (IRS). The roles of endogenous and recombinant resistin were subsequently reversed, using rhRES as the IRS during RES-MSIA quantification. Concurrently, the relative ratio of hRES to rhRES was used as an ancillary technique to rapidly determine the relative concentration of hRES in each of plasma samples. Overall, normal hRES levels determined by RES-MSIA were found to be comparable to those selected and determined by ELISA. With regard to gender, female donor samples were slightly elevated over males. Four single cardiac samples were analyzed and found to have hRES concentrations approximately three times that of the normal. The recombinant-based RES-MSIA is rapid and is amendable to parallel high-throughput robotic processing of resistin related disease cohorts.

Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Publishing Authors By Initials

KA Tubbs,UA Kiernan,EE Niederkofler,D Nedelkov,AL Bieber,RW Nelson,

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Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Analytical chemistry

VOLUME: 78

Page Numbers: 3271-6

Journal Abbreviation: Anal. Chem.

ISSN: 0003-2700

DAY: 15

MONTH: May

YEAR: 2006

Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370536

Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Keywords Mesh Terms:

KEYWORDS: Spectrometry, Mass, Matrix-Assisted Lase

MESH TERMS: methods

Chemical & Substance for Abstract: Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. Information

Substance Name: Resistin

Registry Number: 0

Grant and Affiliation Information for Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling.

AFFILIATION: Intrinsic Bioprobes Inc., 625 South Smith Road, Suite 22, Tempe, AZ 85281, USA. ktubbs@intrinsicbio.com

Country: United States

AGENCY: United States NIEHS

GRANT: N44 ES 35511

ACRONYM: ES

MEDLINETA: Anal Chem

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