Our Mega-ideas for the integration of enzyme or other proteins with nanodevices and incomplex multi-enzyme and protein assemblies hold promise for the future
More detailed information regarding the ANADOLUCA* technology can be found in our patents and publication.
Pub. No.: WO/2011/070402 International Application No.: PCT/IB2009/055707
Publication Date: 16.06.2011 International Filing Date: 11.12.2009
Applicants: SAY, Ridvan [TR/TR]; (TR) Inventors: ERSOZ, Arzu; (TR), et.al,
Abstract: This invention is related to preparation of photosensitive ruthenium based aminoacid monomers and oligomers, aminoacid monomer-protein cross- linking using photo sensitat ion and conjugation on micro and nano-structures by ruthenium-chelate based monomers. Its vast range biotechnolgy applications of multifunctional, biocompatible, stabilE and specific micro and nanobio-conjugates, which will stand-alone or simultaneously enable (i) both purification and determination, (ii) both targeting and imaging and theranostics and (iii) catalysis and determination. The construction and method of preparation is applicable to silica materials, superparamagnetic particles, QDs, CNTs, Ag/ Au nanoparticles and Au surfaces and polymeric materials. The photosensitive aminoacid monomer linkers can react via chemically and biocompatible to a lot of different micro and nano-surface and then to the protein when they act as a single-step cross-linking reaction using irradiation. The photosensitive conjugation based on click biochemistry can be carried out at mild conditions, independent of pH and temperature, without affecting conformation and function of protein.
Microscopy and Microanalysis April 2012 18 : pp 324-330
Copyright © Microscopy Society of America 2012
DOI: http://dx.doi.org/10.1017/S1431927611012840 (About DOI)
Published online: March 2012
Applicants: Rıdvan Say, Gözde Aydoğan Kılıç, Ayça Atılır Özcan, Deniz Hür, Filiz Yılmaz, Adil Denizli and Arzu Ersöz
Abstract: The present study describes the development and use of a new bioconjugate combining targeted quantum dot labeling with an immunoperoxidase method and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1). The new bioconjugate is designed for the examination of samples both under fluorescent and bright-field microscopy at the same time. For this purpose chlorobis(2-2′-bipyridyl) methacryloyl tyrosine-ruthenium(II) and bis (2-2′-bipyridyl) methacryloyltyrosine-methacryloyltryptophan-ruthenium (II) photosensitive monomers and photosensitive poly(Bis (2-2′-bipyridyl)) methacryloyltyrosine-methacryloyltryptophan-ruthenium(II) were synthesized and characterized. The anti-SOD1 antibody and horseradish peroxidase (HRD) conjugated quantum dots were prepared by using this polymer. The anti-SOD1 antibody and HRD conjugated quantum dots were used in labeling and imaging of SOD1 in rat liver sections. Quantum dot particles were observed as a bright fluorescence in their specific binding locations inside the hepatocytes. The HRD-diaminobenzidine reaction product was observed as brown-colored particles at the same locations under bright-field microscopy. Structural details of the tissue sections could be examined at the same time. The conjugation protocol is simple; the bioconjugate is applicable for efficient cell labeling and can be adapted for imaging of other targets in different tissues. Also, the prepared nanobioconjugates have mechanic stability and can be used for a long period.
(Received July 06 2011)
(Accepted November 23 2011)
Abstract: This study presents the development of targeted and antibody cross-linked QDs and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1) on fixed cells and tissues. QD-antibody conjugation was achieved by using our previously invented AmiNoacid (monomer) Decorated and Light Underpining Conjugation Approach (ANADOLUCA) method. In this method, we have used a photosensitive aminoacid monomer having ruthenium complex which is a synthetic and inexpensive material for the preparation of bioconjugates. Its specificity was demonstrated by extracting the active enzyme from rat liver lysate by using the bioconjugate. It provided accurate antibody orientation, high specificity and mechanic stability. The protocol steps for QD-antibody conjugation and specimen preparation were described in detail. The nanobioconjugates were prepared under mild conditions (for example in day light), independent of pH and temperature, without affecting conformation and function of protein. This protocol is simple, inexpensive and can be successfully adapted to detect other targets on different cell types and tissues.