Fresh raw Buffalo milk was frozen (-20℃) overnight. During freezing, much of the lipid content in the milk travels to the top of the milk and forms a bulge. The bulged-out portion is then scraped off the frozen milk and the milk could defrost. Further, to separate the remaining lipid content, the defrosted milk was subjected to refrigerated centrifugation at 10,000 rpm for 30 minutes at 4℃. The whey protein isolate (WPI) was prepared by separating out the major
The protein‐nanoparticles conjugate has been developed as a targeted drug delivery system which selectively and preferentially delivers the therapeutic agents. Here, the alpha‐lactalbumin (ALA) was purified by size exclusion chromatography and confirmed by gel electrophoresis and mass spectrometry (MS). Functionalization of gold nanoparticles (AuNP, ∼29 nm) with ALA was optimized by light scattering and zeta potential measurements. The nanoconjugates (AuNP‐ALA) and ALA showed strong binding and affinity with curcumin and gemcitabine studied using biophysical analysis. The presence of native form of curcumin with ALA was investigated by MS. 8‐anilino‐1‐naphthalenesulfonic acid binding assay showed 95 % surface hydrophilicity of ALA. Circular dichroism and Fourier transform infrared spectroscopy analysis shows increased alpha‐helicity due to the binding of drugs, revealed large increase in non‐covalent interaction. Vesicular membrane interaction with ALA and nanoconjugate exhibited strong interaction at the vesicles surface. The alpha‐lactalbumin‐nanoparticle conjugate in combination with drugs, could be exploited to establish a targeted drug delivery system.
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