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Preparation and Evaluation of Nanoparticles Paclitaxel by Bovine Serum Album Fragments

Author: GaoYuan
Tutor: YuHongRu
School: Liaoning Medical
Course: Biochemistry and Molecular Biology
Keywords: Paclitaxel nanoparticles carrier albumin fragment
CLC: R94
Type: Master's thesis
Year: 2012
Downloads: 95
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Abstract

ObjectiveBy enzymatic hydrolysis of bovine serum albumin, a protein fragment wasscreened from the protein fragments as the new carrier. of paclitaxel nanoparticles,and to examine the feasibility of the new carrier in the physicochemical properties andin vitro release. In order to prepare paclitaxel nanoparticles with less cost.MethodsAlbumin enzymatic hydrolysis experiment was operated by bovine serumalbumin (BSA) and pepsin. The unreacted BSA was removed by columnchromatography. The protein fragments were grouped according to the size ofmolecular weight. Select the appropriate size of the protein fragments as carrier, setBSA as a positive control, in order to prepare paclitaxel nanoparticles by dispersedemulsion-hanging steam. The nanoparticles were observed by transmission electronmicroscopy, and particle size distribution was measured by laser particle size analyzer.Measure the nanoparticle yield by Coomassie brilliant blue method, and measureencapsulation efficiency and drug loading by HPLC method, examine the in vitrorelease by dynamic dialysis method. According to the literature, analysis the data ofsample and reference substance, to examine the feasibility of the protein fragments asa new carrier of paclitaxel nanoparticles.ResultsBSA was digested to a large number of fragments, we got four peaks bycolumn chromatography, SDS-PAGE electrophoresis was operated after the collectionof samples were concentrated. Compared with MARKER, we can estimate themolecular weight of each group as40kD、40kD-31kD、17kD-20kD、17kD.By allergicexperimental verification, the first group and the second group with varying degreesof allergic reactions, the third group and the fourth set of security, including the fourth group only contains one stripe.Selected the fourth group as a candidate carrier and set BSA as positive control,preparation of paclitaxel nanoparticles. Observed by TEM, the appearance of samplegroup and control group were of round and uniform. Average particle size wasmeasured by laser particle sizer as the control group was180.3nm and the samplegroup was150.2nm. After measurement, the average closing rate of the control group(65.100±0.792)%, the average closing rate of the sample group (45.132±0.903)%,the average encapsulation rate of the control group (62.069±0.752)%, with anaverage load dose (4.243±0.090)%, the average encapsulation rate of the samplegroup (50.246±.712), the average drug loading (4.804±0.101)%.From the In vitrorelease profile, we found paclitaxel in the buffer released within one hour, the samplegroup and control group with same profile, cumulative release rates were90.2%and82.4%,with sustained-release effect.ConclusionsThe experiments show that it is feasible to prepare paclitaxel nanoparticles byBSA fragments as carrier. And it is similar with paclitaxel-BSA-nanoparticles atphysical and chemical properties. It is positive for BSA fragments as a carrier forhuman administration. It provides some basis for the expansion of the vector sourceof paclitaxel nanoparticles in theory.

Preparation and Evaluation of Nanoparticles Paclitaxel by Bovine Serum Album Fragments

Abstract

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