Preparation of HIV monoclonal antibody-conjugated pulchellin in order to study its intracellular trafficking pathway in HIV-infected cells by confocal microscopy

M. Sadraeian; F. M. Tsutae; H. H. T. Moreira; A. P. U. Araujo; F. E. G. Guimarães; S. H. Pincus

doi:10.1117/12.2181011

19 June 2015 Preparation of HIV monoclonal antibody-conjugated pulchellin in order to study its intracellular trafficking pathway in HIV-infected cells by confocal microscopy

M. Sadraeian, F. M. Tsutae, H. H. T. Moreira, A. P. U. Araujo, F. E. G. Guimarães, S. H. Pincus

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Proceedings Volume 9531, Biophotonics South America; 953117 (2015) https://doi.org/10.1117/12.2181011
Event: SPIE Biophotonics South America, 2015, Rio de Janeiro, Brazil

Abstract

Pulchellin is a type 2 of ribosome-inactivating proteins isolated from some seeds significantly growing in Brazil. It is a potent agent to inhibit the protein synthesis in cancer cells and also HIV-infected cells. Pulchellin can be conjugated to HIV monoclonal antibodies to specifically target the HIV-infected cells. To analyze the protein synthesis inhibition by Pulchellin, the intracellular localization of the immunoconjugate should be compared to Pulchellin. In this case, the intracellular trafficking of this protein in cells can be determined by confocal microscopy. In our study, we utilized Pulchellin to construct HIV monoclonal antibody-conjugated Pulchellin A chain in order to target HIV-infected lymphocyte cells. Afterward the conjugation was labeled with the superior Alexa Fluor 488 dye. As a subsequent step, we are interested in studying the intracellular trafficking pathway of this novel conjugation in HIV-infected cells by confocal microscopy. Moreover, possible quantitative methods for fluorescent labeling of the immunoconjugate during confocal microscopy will be investigated.

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M. Sadraeian, F. M. Tsutae, H. H. T. Moreira, A. P. U. Araujo, F. E. G. Guimarães, and S. H. Pincus "Preparation of HIV monoclonal antibody-conjugated pulchellin in order to study its intracellular trafficking pathway in HIV-infected cells by confocal microscopy", Proc. SPIE 9531, Biophotonics South America, 953117 (19 June 2015); https://doi.org/10.1117/12.2181011

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KEYWORDS

Proteins

Confocal microscopy

Monoclonal antibodies

Molecules

Picture Archiving and Communication System

Fluorescence correlation spectroscopy

Cancer

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