Development and characterization of new rat monoclonal antibodies for procalcitonin

Analytical and Bioanalytical Chemistry
Petra M KrämerElisabeth Kremmer

Abstract

The development of selective and sensitive biological recognition elements, e.g., antibodies, for the detection of relevant blood markers is a great challenge in the field of biosensors. In this context, five new rat monoclonal antibodies (mAbs) for procalcitonin (PCT), a marker for bacterial infection and sepsis, were developed and characterized. One mAb, PROC1 3G3, was used as capture antibody. Four mAbs, PROC4 6C6, PROC4 6B2, PROC4 1G3, and PROC4 1D6, were used as detection mAbs, either as Protein G-purified or as biotinylated mAbs. A surface plasmon resonance (SPR) biosensor was used to characterize the antigen-antibody biomolecular interactions. The capture mAb (PROC1 3G3) has an equilibrium dissociation constant (K (D)) of 3.42 x 10(-8) M. All four detection mAbs (PROC4 6C6, PROC4 6B2, PROC4 1G3, and PROC4 1D6) are of high affinity (K (A) = 2.81-6.11 x 10(8) M(-1); K (D) = 1.64-3.56 x 10(-9) M) and have moderate dissociation rate constants (k (d) = 1.70-2.40 x 10(-3) s(-1)). Four different sandwich enzyme-linked immunosorbent assays (ELISAs) with standards of human recombinant (hr) PCT, using PROC1 3G3 as capture mAb and PROC4 mAbs as detection mAbs, respectively, led to highly specific determinations of PCT without cross...Continue Reading

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Citations

Dec 20, 2008·Analytical and Bioanalytical Chemistry·F BaldiniC Trono
Oct 19, 2011·Analytical and Bioanalytical Chemistry·Elisabeth KremmerPetra M Krämer
Dec 4, 2009·Nucleic Acids Research·Yusuke MiyachiAkihiko Kondo
Feb 11, 2015·Clinica Chimica Acta; International Journal of Clinical Chemistry·Haiying WangJufang Wang
Dec 18, 2009·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Aug 30, 2020·Animals : an Open Access Journal From MDPI·Federica BattagliaLuigi Intorre

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