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International Journal of Latest Research in Science and Technology

DOI:10.29111/ijlrst   ISRA Impact Factor:3.35

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INFLUENCE OF DIFFERENT EXCIPIENTS AND PROTEIN CHARACTERISTICS ON THEIR SUSCEPTIBILITY TO MICROENCAPSULATION PROCESS

Research Paper Open Access

International Journal of Latest Research in Science and Technology Vol.3 Issue 4, pp 31-36,Year 2014

INFLUENCE OF DIFFERENT EXCIPIENTS AND PROTEIN CHARACTERISTICS ON THEIR SUSCEPTIBILITY TO MICROENCAPSULATION PROCESS

Reginaldo Almeida da Trindade,Ant?nio Carlos Melo Lima-Filho,Mouzarllem Barros dos Reis, Lucas Rodrigues de Carvalho,Paulo Henrique de Holanda Veloso

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Received : 30 July 2014; Accepted : 09 August 2014 ; Published : 31 August 2014

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Abstract

This study evaluated the effects of microencapsulation process on different proteins. Insulin, albumin and casein were used as model proteins. Three methodologies to determine protein concentrations were compared: λ 280 nm, Lowry and Bradford colorimetric methods. Proteins were submitted to the first phase of double emulsion method, which is commonly used to produce polymeric delivery systems. Different excipient solutions were tested: NaCl, KSCN, MgCl2 and NaH2PO4 all at 50 mM, PBS and H2O were used as controls. Structural analysis of proteins was performed by SDS-PAGE and intrinsic fluorescence intensities. Results showed direct reading at 280 nm the most suitable for quantitative measures of the three proteins presenting linear coefficient (r2) of 0.9849, 0.9992 and 0.9995 for insulin, albumin and casein, respectively. The excipient solution that allowed greater recovery of proteins was PBS, which recovered 45%, 44% and 90% of albumin, insulin and casein, respectively. For albumin, KSCN solution also enabled recovering 47% of processed protein. Electrophoretic pattern did not revealed fragmentation or aggregation of the three analyzed proteins. Structural analysis of proteins by measuring of intrinsic fluorescence has shown: PBS and NaH2PO4 solutions effectively maintained the integrity of protein structures for albumin, KSCN and MgCl2 solutions for insulin, and PBS solution for casein. Taken all these results together, it was possible to conclude that phosphate-based solutions are best excipients to protect proteins with higher molecular weights or size, and chaotropic salts, such as MgCl2, are the best one to low-molecular weight peptides as insulin.

Key Words   
Microspheres; PLGA; Protein stability; Protein encapsulation.

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To cite this article

Reginaldo Almeida da Trindade,Ant?nio Carlos Melo Lima-Filho,Mouzarllem Barros dos Reis, Lucas Rodrigues de Carvalho,Paulo Henrique de Holanda Veloso , " Influence Of Different Excipients And Protein Characteristics On Their Susceptibility To Microencapsulation Process ", International Journal of Latest Research in Science and Technology . Vol. 3, Issue 4, pp 31-36 , 2014

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