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A multi-functional polyhydroxybutyrate nanoparticle for theranostic applications
Kwon, Hee-Su,Jung, Sung-Geun,Kim, Hae-Yeong,Parker, Stephanie A.,Batt, Carl A.,Kim, Young-Rok The Royal Society of Chemistry 2014 Journal of Materials Chemistry B Vol.2 No.25
<P>Biopolymer-based multi-functional nanoparticles have been developed through a one-step enzymatic polymerization reaction using engineered polyhydroxyalkanoate (PHA) synthase fused with green fluorescent protein (GFP) and a single chain variable fragment antibody (A33scFv) specific to colon cancer. PHA synthase possesses unique catalytic characteristics, namely covalent catalysis, by which the synthesized polyhydroxybutyrate chain remains covalently attached to the enzyme. The amphiphilic nature of the resulting protein-polymer hybrid gives rise to spontaneous self-assembly into a micellar structure with GFP and A33scFv displayed on the surface (AGPHB nanoparticle). A model compound, Nile red, was loaded into the hydrophobic core of the AGPHB nanoparticle during the polymerization and self-assembly process. The specificity of the fluorescent multi-functional AGPHB nanoparticle towards the colon cancer cell lines SW1222 (A33+) and HT29 (A33−) was confirmed and analysed quantitatively <I>in vitro</I>. This new biological approach provides a simple means of producing nanocarriers with a range of surface functionality and the sizes desired for imaging and targeted drug delivery.</P>
Expression of Mature Thaumatin in Escherichia coli
함영태,Batt, Carl A. 中央大學校 遺傳工學硏究所 1991 遺傳工學硏究論集 Vol.4 No.1
Thaumatin, a sweet-tasting plant protein, is initially produced as a precursor, preprothaumatin. The presignal sequence at aminoterminal end containing 22 mostly hydrophobic amino acid residues and the prosignal sequence at carboxy-terminal end carrying 6 mainly acidic amino acid residues of preprothaumatin II were removed by using a synthetic oligonucleotide and site-specific mutagenesis to produce mature thaumatin in E. coli. The presignal sequence was replaced by a 34mer oligonucleotide carrying a Shine-Dalgarno sequence and a translational initiation codon. The first amino acid of prosignal sequence, leu-230, was changed to an ochre stop codon TAA. The met-thaumatin gene in the plasmid pUTH-3 carrying two copies of the lac UV5 promoters was expressed in E. coli. A trace amount of an immunoreactive 22 kDa protein of a molecular weight similar to mature thaumatin was observed in E. coli SVS-3 [pUTH-3].
Multiplexed Protein Patterns on a Photosensitive Hydrophilic Polymer Matrix
Bhatnagar, Parijat,Malliaras, George G.,Kim, Il,Batt, Carl A. WILEY-VCH Verlag 2010 Advanced Materials Vol.22 No.11
<B>Graphic Abstract</B> <P>A photosensitive hydrogel film containing a photocleavable protecting group is attached to silicon oxide. Projection photolithography is used to generate carboxylic groups in selective patterns on this film, to which proteins are coupled through their primary amine groups. The carboxylic groups on the proteins are chemically protected and the process is repeated to sequentially pattern additional proteins on the hydrogel surface. <img src='wiley_img_2010/09359648-2010-22-11-ADMA200903255-content.gif' alt='wiley_img_2010/09359648-2010-22-11-ADMA200903255-content'> </P>
Encapsulation of Single Small Gold Nanoparticles by Diblock Copolymers
Chen, Hong Y.,Abraham, Sinoj,Mendenhall, Juana,Delamarre, Soazig C.,Smith, Kahli,Kim, Il,Batt, Carl A. WILEY-VCH Verlag 2008 CHEMPHYSCHEM -WEINHEIM- Vol.9 No.3
<B>Graphic Abstract</B> <P>Hairy micelles are obtained by using amphiphilic diblock copolymers with long hydrophilic blocks which favor single encapsulation of Au nanoparticles (ca. 5 nm, see figure). Antibody molecules can be attached to the hydrogel layers of the encapsulated Au nanoparticles to give nanoparticle/biomolecule conjugates. <img src='wiley_img/14394235-2008-9-3-CPHC200700598-content.gif' alt='wiley_img/14394235-2008-9-3-CPHC200700598-content'> </P>