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An enzyme-linked immunosorbent assay...

Rahman, Moshiur.

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An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine./
Author:	Rahman, Moshiur.
Description:	45 p.
Notes:	Source: Masters Abstracts International, Volume: 48-02, page: 1047.
Contained By:	Masters Abstracts International48-02.
Subject:	Chemistry, Biochemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=1472520
ISBN:	9781109493269

An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine.
Rahman, Moshiur.

An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine. - 45 p.

Source: Masters Abstracts International, Volume: 48-02, page: 1047.

Thesis (M.S.)--Stephen F. Austin State University, 2009.

Quantitating elastin in tissues or determining the loss of elastin from tissues during various disease or surgical procedures has become an important tool in basic research and clinical environments. In recent years desmosine and isodesmosine, crosslinking amino acids unique to elastin, have been used for this purpose. It was the goal of this thesis to design a rapid and accurate desmosine and isodesmosine ELISA that could replace the RIA as a routine means of making these measurements. Experiments were performed that optimize the desmosine-ovalbumin conjugate for coating microtiterplate wells. Two dimensional serial dilutions of the primary and secondary antibodies established the best assay conditions for both the desmosine and isodesmosine ELISA. Desmosine values from mouse lung hydrolysates obtained by ELISA were in good agreement with values obtained by RIA from the same hydrolysates. Cross-reactivity between desmosine and isodesmosine ranged from 14--23%. Urine desmosine values were significantly higher when assayed by ELISA compared to the RIA and that makes the use of the ELISA for urine desmosine or isodesmosine content questionable. The assay when used for tissue hydrolysates was simple, precise, reproducible and could be performed within 4hr.

ISBN: 9781109493269Subjects--Topical Terms:

1017722
Chemistry, Biochemistry.

An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine.
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020 $a 9781109493269
035 $a (MiAaPQ)AAI1472520
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100 1 $a Rahman, Moshiur. $3 2097657
245 1 3 $a An enzyme-linked immunosorbent assay (ELISA) to quantitate desmosine and isodesmosine.
300 $a 45 p.
500 $a Source: Masters Abstracts International, Volume: 48-02, page: 1047.
500 $a Advisers: Barry Starcher; Robert Stewart.
502 $a Thesis (M.S.)--Stephen F. Austin State University, 2009.
520 $a Quantitating elastin in tissues or determining the loss of elastin from tissues during various disease or surgical procedures has become an important tool in basic research and clinical environments. In recent years desmosine and isodesmosine, crosslinking amino acids unique to elastin, have been used for this purpose. It was the goal of this thesis to design a rapid and accurate desmosine and isodesmosine ELISA that could replace the RIA as a routine means of making these measurements. Experiments were performed that optimize the desmosine-ovalbumin conjugate for coating microtiterplate wells. Two dimensional serial dilutions of the primary and secondary antibodies established the best assay conditions for both the desmosine and isodesmosine ELISA. Desmosine values from mouse lung hydrolysates obtained by ELISA were in good agreement with values obtained by RIA from the same hydrolysates. Cross-reactivity between desmosine and isodesmosine ranged from 14--23%. Urine desmosine values were significantly higher when assayed by ELISA compared to the RIA and that makes the use of the ELISA for urine desmosine or isodesmosine content questionable. The assay when used for tissue hydrolysates was simple, precise, reproducible and could be performed within 4hr.
520 $a The ideal assay should be sensitive enough to measure low levels of desmosine found in urine, yet broad enough to measure samples very high in elastin content like vascular tissues, without unnecessary dilutions. The assay should be robust in terms of reagent stability and applicable to samples of variable pH and salt concentrations. The assay should be rapid and suited for multi sample analysis and robotic machines. Most importantly the assay needs to be precise and reproducible with low intra and inter sample variability.
520 $a In conclusion, an ELISA was developed for desmosine and isodesmosine that performed well for tissue hydrolysates, yielding values equivalent to those found in the literature or with the same samples run by RIA. The ELISA was of limited use with urine so far due to apparent interfering substances.
590 $a School code: 6340.
650 4 $a Chemistry, Biochemistry. $3 1017722
650 4 $a Chemistry, Analytical. $3 586156
690 $a 0487
690 $a 0486
710 2 $a Stephen F. Austin State University. $3 1020624
773 0 $t Masters Abstracts International $g 48-02.
790 $a 6340
791 $a M.S.
792 $a 2009
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=1472520