Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing

John S. Mitchell, Yinqiu Wu, Christian J. Cook, Lyndsay Main

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Thioether-linked 3-mercaptopropionic acid derivatives of 17β-estradiol and estrone were formed at the A-ring 4-position of the steroids by substitution of their 4-bromo analogues. The carboxylic acid terminal was used to link to an oligoethylene glycol (OEG) chain of 15-atoms in length. The OEG derivative of 17β-estradiol was then in situ immobilized on a carboxymethylated dextran-coated gold sensor surface used to detect refractive index changes upon protein binding to the surface by surface plasmon propagation in a BIAcore surface plasmon resonance (SPR) instrument. Two other estradiol-OEG derivatives with Mannich reaction linkage at the 2-position and hemisuccinate linkage at the 3-position were also immobilized on the sensor surfaces for comparison. Binding performance between these immobilized different positional conjugates and monoclonal anti-estradiol antibody, raised from a 6-position conjugate, clearly demonstrated that both 2- and 4-conjugates, not conjugated through existing functional groups, gave strong antibody bindings, whereas the 3-conjugate through an existing functional group (3-OH) gave very little binding (2% compared to the 2-conjugate). Both 2- and 4-position conjugates were then applied in a highly sensitive estradiol SPR immunoassay with secondary antibody mediated signal enhancement that gave up to a 9.5-fold signal enhancement of primary antibody binding, and a detection limit of 25 pg/mL was achieved for a rapid and convenient flow-through immunoassay of estradiol.

Original languageEnglish
Pages (from-to)618-631
Number of pages14
JournalSteroids
Volume71
Issue number7
DOIs
Publication statusPublished - Jul 2006
Externally publishedYes

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Surface Plasmon Resonance
Surface plasmon resonance
Estradiol
Estrogens
Glycols
Antibodies
Derivatives
Immunoassay
Functional groups
3-Mercaptopropionic Acid
Refractometry
Estrone
Sensors
Sulfides
Carboxylic Acids
Dextrans
Protein Binding
Gold
Limit of Detection
Anti-Idiotypic Antibodies

Cite this

Mitchell, John S. ; Wu, Yinqiu ; Cook, Christian J. ; Main, Lyndsay. / Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing. In: Steroids. 2006 ; Vol. 71, No. 7. pp. 618-631.
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Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing. / Mitchell, John S.; Wu, Yinqiu; Cook, Christian J.; Main, Lyndsay.

In: Steroids, Vol. 71, No. 7, 07.2006, p. 618-631.

Research output: Contribution to journalArticle

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