Fingerprint-Imprinted Polymer: Rational Selection of Peptide Epitope Templates for the Determination of Proteins by Molecularly Imprinted Polymers

The pool of peptides composing a protein allows for its distinctive identification in a process named fingerprint (FP) analysis. Here, the FP concept is used to develop a method for the rational preparation of molecularly imprinted polymers (MIPs) for protein recognition. The fingerprint imprinting...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2012-05, Vol.84 (9), p.4036-4041
Main Authors: Bossi, Alessandra M, Sharma, Piyush S, Montana, Luca, Zoccatelli, Gianni, Laub, Orgad, Levi, Raphael
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analytical chemistry
Cardiovascular Diseases - diagnosis
Chemistry
Exact sciences and technology
Humans
Molecular Imprinting - methods
Molecules
Natriuretic Peptide, Brain - blood
Natriuretic Peptide, Brain - isolation & purification
Natriuretic Peptide, Brain - metabolism
Peptide Fragments - blood
Peptide Fragments - isolation & purification
Peptide Fragments - metabolism
Peptides
Peptides - chemistry
Peptides - metabolism
Polymers
Polymers - chemistry
Polymers - metabolism
Protein Binding
Proteins
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Summary:The pool of peptides composing a protein allows for its distinctive identification in a process named fingerprint (FP) analysis. Here, the FP concept is used to develop a method for the rational preparation of molecularly imprinted polymers (MIPs) for protein recognition. The fingerprint imprinting (FIP) is based on the following: (1) the in silico cleavage of the protein sequence of interest with specific agents; (2) the screening of all the peptide sequences generated against the UniProtKB database in order to allow for the rational selection of distinctive and unique peptides (named as epitopes) of the target protein; (3) the selected epitopes are synthesized and used as templates for the molecular imprinting process. To prove the principle, NT-proBNP, a marker of the risk of cardiovascular events, was chosen as an example. The in silico analysis of the NT-proBNP sequence allowed us to individuate the peptide candidates, which were next used as templates for the preparation of NT-pro-BNP-specific FIPs and tested for their ability to bind the NT-proBNP peptides in complex samples. Results indicated an imprinting factor, IF, of ∼10, a binding capacity of 0.5–2 mg/g, and the ability to rebind 40% of the template in a complex sample, composed of the whole digests of NT-proBNP.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac203422r