MIP_presentation - O-NH 3 CH 2-CH 2-CH 2-CH 3 OH N NH 2 Cl...

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Applications of Molecular Imprinting Technology to Immunoassays Roger L. Bertholf, Ph.D. Associate Professor of Pathology University of Florida Health Science Center/Jacksonville
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Early theories of antibody formation Paul Ehrlich (1854-1915) proposed that antigen combined with pre-existing side-chains on cell surfaces. Ehrlich’s theory was the basis for the “genetic theory” of antibody specificity.
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The “Template” theory of antibody formation Karl Landsteiner (1868-1943) was most famous for his discovery of the A/B/O blood groups and the Rh factor. Established that antigenic specificity was based on recognition of specific molecular structures; he called these “haptens”; formed the basis for the “template” theory of antibody formation.
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History of molecular imprinting Linus Pauling (1901-1994) first suggested the possibility of artificial antibodies in 1940 Imparted antigen specificity on native globulin by denaturation and incubation with antigen .
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The birth of immunoassay Rosalyn Yalow (1921-) and Solomon Berson described the first radioimmunoassay in 1957.
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Fundamentals of antigen/antibody interaction
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Unformatted text preview: O-NH 3 + CH 2-CH 2-CH 2-CH 3 OH N NH 2 Cl Molecular imprinting (Step 1) N N O N NH O H 3 C CH 3 N N O N NH O H 3 C CH 3 Methacrylic acid + Porogen Molecular imprinting (Step 2) N N O N NH O H 3 C CH 3 N N O N NH O H 3 C CH 3 Molecular imprinting (Step 3) N N O N NH O H 3 C CH 3 N N O N NH O H 3 C CH 3 Cross-linking monomer Initiating reagent Molecular imprinting (Step 4) Comparison of MIPs and antibodies • In vivo preparation • Limited stability • Variable specificity • General applicability • In vitro preparation • Unlimited stability • Predictable specificity • Limited applicability Antibodies MIPs Immunoassays using MIPs • Therapeutic Drugs: Theophylline, Diazepam, Morphine, Propranolol, Yohimbine ( α 2-adrenoceptor antagonist) • Hormones: Cortisol, Corticosterone • Neuropeptides: Leu 5-enkephalin • Other: Atrazine, Methyl-α-glucoside Aptamers 10 14-10 15 random sequences Target Oligonucleotide-Target complex Unbound oligonucleotides Aptamer candidates PCR New oligonucleotide library + Target...
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This note was uploaded on 09/18/2011 for the course MED 6566 taught by Professor Staff during the Summer '11 term at University of Florida.

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MIP_presentation - O-NH 3 CH 2-CH 2-CH 2-CH 3 OH N NH 2 Cl...

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