INTEGRATION OF NANOPARTICLES WITH PROTEIN MICROARRAYS

A novel method for immobilization of proteins via entrapment of magnetic nanoparticles through epoxy cross-linking

Projector-based display systems have been used in area of computer interaction as Ad-hoc interface in recent time. The mobile hand-held projectors are becoming more popular. Many human centric user interfaces with the human wearable computer are being developed. Most of such system uses daily objects for projection and the interaction. But most of ignores the fact that these object surfaces are not planar. Hence such interfaces suffers from the distortion due to non-planar projection surface. Besides this projection quality also suffers from the radiometric distortion as well. Further more the interaction proposed with such interfaces bound to the planar surface only. Hence this paper is targeted to address the geometric distortion free projection of and interaction with such interfaces on non planar surfaces. Kinect is used as depth sensor for 3D scenario acquisition. We use image-per warping to mesh from Kinect. We use colored fingertip gloves for interaction. Here our system aims any day to day object surface for distortion free projection such as human body, curved wall, room corners, curtain's and many more objects.

Chapter 10 - Single Molecule Microarray Analysis

The present communication addresses the reflection and transmission analysis of a two-dimensional waveguide structure with planar and non-planar surfaces. In the fields of physics, engineering and applied mathematics, research is being carried out to control or reduce unwanted and harmful noise to an acceptable level. Our aim is to solve the boundary value problem developed to simulate the transmission and reflection of a bifurcated waveguide structure in the channels or ducts that have abrupt changes in the height together with planar and non-planar surfaces. Furthermore, the eigenfunctions are linearly independent and non-orthogonal. To solve non-orthogonal eigenfunctions, a generalized orthogonality relation has been developed. Solution is obtained using mode matching technique (MMT) along with generalized orthogonal relations. In the last section, solution is viewed graphically in the inner and outer ducts or channels for different frequencies and height. The expressions for energy distribution in different duct regions are obtained and sketched subject to the various properties of the waveguide material. The results are well supported for a number of mathematical and physical reasons.

Miniaturization and multiplexing are two areas that need improvement in immunoassays. However, it is unclear how associated changes in size, substrate, binding capacity, protein concentration, and other parameters affect sensitivity and quantitative range. In the study here we compared a new protein microarray substrate and system (Zeta-Grip, Miragene Inc., Anaheim, CA) with enzyme-linked immunosorbent assay (ELISA) for the application of immunoassays. We found that the protein microarray system performed better than ELISA in both sensitivity and quantitative range for use in immunoassay.

An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.

Substantial progress in biochip technologies has established an efficient and reliable platform for advanced biological and biomedical applications. In particular, the use of protein biochips in high‐throughput screens provides high content information. We briefly introduce here recent developments in protein biochip preparation with a special focus on our own work on new methods for protein immobilization and protein microarray fabrication, including the application of the Diels‐Alder reaction, Staudinger ligation, ‘click’ sulfonamide formation, and the photochemical thiol‐ene reaction. These chemical methods allow for oriented, site‐specific protein conjugation on solid surfaces with high sensitivity and specificity under mild, aqueous conditions.magnified image

Receptor chromatography is an efficient analytical technique that combines the high separation ability of chromatography with the high specificity of receptors for drug recognition. In addition, this technique offers the advantages of active recognition, online separation, and convenient multidimensional target tracking. This strategy allows target active ingredients in complex systems, such as traditional Chinese medicines, to be efficiently screened and accurately identified. Furthermore, the interactions between ligands and immobilized proteins can be studied. To avoid a loss in function, receptor chromatography requires efficient, mild, and simple immobilization methods that do not damage the structure of the immobilized receptors. Improvements in the activity, stability, and ligand-recognition specificity of immobilized functional proteins can be achieved by selecting appropriate immobilization conditions. Notably, the protein immobilization method is not only closely related to the recognition ability of receptor chromatography but also determines the accuracy of the technique. Common methods for immobilizing functional proteins include physical adsorption, chemical reactions, biological affinity reactions, and click chemistry. Despite being easy to operate under mild reaction conditions, these methods have shortcomings, including poor reaction specificity and the necessity of using high-purity functional proteins to prepare chromatography columns. Maintaining the high activity of immobilized receptors and ensuring excellent identification and separation abilities are key challenges in the further development of receptor chromatography. In this work, these issues were addressed by introducing a specific bioorthogonal reaction involving haloalkane dehalogenase (Halo) and 6-chlorohexanoic acid for the immobilization of the α1A-adrenergic receptor (α1A-AR). Specifically, Halo-α1A-AR was immobilized on the surface of 6-chlorohexanoic acid-modified aminopropyl silica gel in one step. The stationary phase with immobilized Halo-α1A-AR was characterized using scanning electron microscopy. Moreover, the activity of the Halo-α1A-AR chromatographic column was evaluated using specific ligands (terazosin hydrochloride, phentolamine mesylate, tamsulosin hydrochloride, and urapidil) and nonspecific ligands (yohimbe and metoprolol) for α1A-AR. Halo-α1A-AR was successfully immobilized on the silica gel surface with good stability over 30 days, and the Halo-α1A-AR chromatographic column exhibited good ligand-recognition activity. The nonlinear chromatography results indicated that prazosin hydrochloride, terazosin hydrochloride, and urapidil interacted with immobilized Halo-α1A-AR through one type of binding site, with association constants of 3.85×105, 5.00×105, and 5.90×105L/mol, respectively. In contrast, phentolamine mesylate and tamsulosin hydrochloride interacted with immobilized Halo-α1A-AR through two types of binding site. The association constants with the high- and low-affinity binding sites were 3.12×106 and 6.01×105L/mol, respectively, for phentolamine mesylate and 9.98×105 and 0.21×105L/mol, respectively, for tamsulosin hydrochloride. Compared with the traditional carbonyldiimidazole method, the immobilization method developed in this work did not require receptor purification and thus minimized the loss of receptor activity. The affinity constants obtained with immobilized Halo-α1A-AR were consistent with the values determined for receptor-ligand binding in solution, indicating that the Halo-α1A-AR chromatography column is suitable for studying drug-protein interactions. This approach also provides a foundation for the efficient screening and accurate determination of target active ingredients in complex systems.

Objective To screen the proteins that are related to the sensitivity or resistance of neoadjuvant chemotherapy in breast cancer and to provide molecular basis on the prediction model.Methods 10 patients were included in this study,who received neo-adjuvant taxane and anthracyclinebased chemotherapy,total protein was extracted from core needle biopsy samples of which 5 were sensitive to neo-adjuvant chemotherapy and 5 were resistant to neo-adjuvant chemotherapy.The sensitivity criterion was based on Miller & Payne grading,grade 4 and grade 5 were considered as sensitive,grade 1 and grade 2 were considered as resistant.Protein profiling was analyzed by antibody microarray,then screening of sensitive and resistant related proteins and clustering analysis were conducted.Results Core needle biopsy can provide sufficient protein to conduct the antibody microarray analysis,61 differentially expressed proteins were identified,10 patients were grouped as sensitive or resistant to neo-adjuvant chemotherapy by clustering analysis with an accuracy of 100%.Conclusion Molecular subtype of breast cancer based on proteomic analysis has a potential value to predict the sensitivity to neo-adjuvant chemotherapy. Key words: Breast neopalsms; Protein array analysis; Antineoplastic combined chemotherapy protocols

A mass production method of label-free protein microarray integrated with micro-well structures for the use of miniaturized multi-parallel scanning system was investigated. The geometrical parameters of biosensing structure were designed by rigorous coupled wave analysis simulation, and micro-well structures were designed considering the detection and material delivery system. The protein microarray with micro-well structures was fabricated by one-step UV nanoimprinting process using an electroformed multi-scale metallic stamp. Finally, microarray scanning was achieved using optical modulation without applying any motorized system and the feasibility of proposed protein microarray and scanning system was demonstrated by verifying the bio-molecular interactions.

Clinical OMICsVol. 8, No. 4 NewsPowering Diagnostics for Infectious Disease: The COVID-19 pandemic saw a rush of companies developing diagnostics for the infectious disease that should benefit the detection of other diseasesMalorye Allison BrancaContributing EditorMalorye Allison BrancaContributing EditorSearch for more papers by this authorPublished Online:26 Aug 2021https://doi.org/10.1089/clinomi.08.04.24AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetails Volume 8Issue 4Jul 2021 InformationCopyright © GEN PublishingTo cite this article:Malorye Allison BrancaContributing Editor.Powering Diagnostics for Infectious Disease: The COVID-19 pandemic saw a rush of companies developing diagnostics for the infectious disease that should benefit the detection of other diseases.Clinical OMICs.Jul 2021.36-39.http://doi.org/10.1089/clinomi.08.04.24Published in Volume: 8 Issue 4: August 26, 2021PDF download

Irregularities at the {001} surface of MgO: Topography and other aspects

Covalent immobilization of protein monolayers for biosensor applications

Plasma treatment of paper for protein immobilization on paper-based chemiluminescence immunodevice

Progress in protein and antibody microarray technology