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Design and Construction of Novel Molecular Conjugates for Signal Amplification (II): Use of Multivalent Polystyrene Microparticles and Lysine Peptide Chains to Generate Immunoglobulin-Horseradish Peroxidase Conjugates

Design and Construction of Novel Molecular Conjugates for Signal Amplification (II): Use of Multivalent Polystyrene Microparticles and Lysine Peptide Chains to Generate Immunoglobulin-Horseradish Peroxidase Conjugates

Subhash Dhawan

Peptides. 2002 Dec;23(12):2099-110.

PMID: 12535688

Abstract:

Spherical polystyrene microparticles expressing a large number of highly reactive functional groups were chemically engineered to generate antibody-enzyme conjugates as novel signal amplification systems. Chemically modified goat anti-human IgG and horseradish peroxidase (HRP) were combined in a 1:5 ratio and attached to 0.44 microm streptavidin microparticles or N-succinimidyl-S-acetylthioacetate (SATA)-activated 0.29 microm amino microparticles with highly reactive free sulfhydryl groups on their surface. The numbers of HRP molecules/microparticle were further increased by coupling HRP to primary amines on N-terminal biotinylated or bromoacetylated polypeptides containing 20 lysine residues prior to conjugation with streptavidin or sulfhydryl groups-containing microparticles. The antibody-poly-HRP immunoconjugates contained an estimated number of 10(5)HRP/streptavidin microparticle and 10(6)HRP/amino microparticle, respectively. These microparticle immunoconjugates efficiently bound to plasma anti-HIV-1 antibodies that had been captured by HIV antigens on 5 microm carboxyl magnetic microparticles and, upon reaction with orthophenyldiamine substrate, produced a detection signal with 5-8 times more sensitivity as compared to conventional HRP-conjugated goat anti-human IgG. The signal amplification technique by microparticle immunoconjugates may provide potentially novel tools for the development of highly sensitive diagnostic systems.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42414006	Anti-Coronin 1/TACO antibody produced in goat			Price

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