Tag: Porous

Multiplexed ONCYTE Protein microarrays Used for Infectious Disease Immunogen Discovery

First, in an interesting Shistotosomiasis study, Gaze, et al, employ protein microarrays in a longitudinal immunomics study for vaccine antigen discovery (1).  Schistosomiasis is a pervasive disease affecting 200 million people a year and traditional drug treatments are ineffective due to re-infection, drug expense, and parasite drug resistance. Non-sterile immunity does develop over long periods of … Continued

A Comprehensive Technology Survey of Protein Microarray Platforms, Techniques and Detection Methods

A comprehensive assessment of multiplex immunoassay platforms and detection methods is summarized in an interesting 2011 article by Chandra, et al, in Expert Review Proteomics (1). The article, “Protein Microarrays and Novel Detection Platforms” provides interesting information for several reasons.First, it presents an excellent description of the emerging discovery and clinical need for high throughput … Continued

Sensitive and Accurate Multi-Color RPPA Using Grace Bio-Labs ArrayCAM Protein Microarray System

Introduction Profiling sets of multiple protein markers and signaling pathways is becoming the standard method for biomarker discovery, accurate disease diagnosis, and monitoring of therapeutic response. Low-cost, high throughput multiplexed assay platforms that can provide specific, sensitive, and reproducible measurements, featuring reliable and robust assay controls along with easy to use methodologies will yield increased … Continued

Improved Classification of Influenza Infection, Future Susceptibility and Pandemic Epidmiology using ONCYTE Porous Nitrocellulose Film Microarrays

An interesting paper furthering the expansion of infectious disease serological applications using ONCYTE porous nitrocellulose film slides was described by researchers out of the Netherlands for classifying influenza infections and susceptibility to future infections. Microarrays printed on ONCYTE AVID film slides used purified recombinant influenza virus hemagglutinin antigens (HA1) from various subtypes (H1, H2, H3, … Continued

Targeted Malarial Serological Diagnostics using ONCYTE Porous Nitrocellulose Film Microarray

Targeted diagnostic serological screening using ONCYTE porous nitrocellulose (PNC) microarrays is currently greatly expanding. Interesting research in the Felgner laboratory at UC Irvine has centered on identifying and monitoring P. falciparum antigens involved in malaria infection using subsets of its viral proteome produced via a recombinant expression system and arraying on PNC. Recently, in collaboration … Continued

Review of Mujawar et al Analytical Chemistry – Distribution of Biomolecules in Porous Nitrocellulose Films

An interesting paper in (Analytical Chemistry) describes the use of confocal fluorescence section imaging to analyze protein distribution in microarray spots on nitrocellulose film slides. By scanning and imaging through the Z dimension of a printed spot of fluor labeled protein, a 3-dimensional quantitation of fluorophore density in the films was obtained, showing that commercial … Continued

Multiplex Microarrays: The Next Generation ELISA

The hallmark of chronic diseases is the production of highly specific autoantibodies (1,2).  These autoantibodies are increasingly measured to guide clinical decision making.    The goal of these measurements is to define intracellular signal pathway activation profiles. These profiles can be integrated with clinical information to generate “biosignatures” for individual patients, the goal of which is personalized … Continued

Sensitive Detection of Total Protein on RPPA using Fast Green

Reverse phase protein arrays (RPPA) are designed for sensitive quantitation of a target protein in a very small sample of tissue lysate, and multiplex analysis of many samples in one experiment. This technology holds great promise for the identification and analysis of critical biomarkers for disease states in patient samples and for high-throughput drug screening … Continued

A Systems Approach to Quantitative Multiplexed RPPA

A SYSTEMS APPROACH TO QUANTITATIVE MULTIPLEXED RPPA’S Presented by: Michael A. Shultz, PhD  RPPA Workshop in Kobe, Japan November, 2013 Porous nitrocellulose film (PNC) is the substrate of choice for RPPA largely because of its extraordinarily high protein binding capacity, which can exceed non-porous planar counterparts by 500X.

High Capacity Porous Nitrocellulose Microarray Substrates Enable Whole Proteome Serology Screening Assays for Antigenicity Profiling of Infectious Disease Agents

Insight into the dynamics of pathogenic infectivity has been greatly enhanced by studies using whole proteome microarray technology [1]. Techniques allowing high throughput production of recombinant proteins representing comprehensive antigenic potential of pathogenic bacteria and viruses allow examination of the degrees to which specific antigenic proteins are involved in human serological response.The true value of … Continued

Beating Background Fluorescence on Nitrocellulose Film-Slides

Near Infrared Detection with Porous Nitrocellulose Films Improves Sensitivity for Protein Arrays. Despite some clear performance advantages of porous nitrocellulose Film-Slides for protein arrays, one objection to their use is the inherent fluorescence background detected on most scanners when using popular organic dyes with visible emission wavelengths (500-650 nm). However, fluorescence detection in near infrared … Continued

Optimized Blocking Of Porous Nitrocellulose Films For Sensitive Protein Microarrays

Michael A. Shultz, Aki Ohdera, Jason MacManiman, Charles M. McGrath. Grace Bio-Labs, Inc.   ABSTRACT Super G Blocking Buffer was developed to produce low backgrounds in assays using nitrocellulose film-slides.  Using this blocking reagent in combination with ONCYTE® SuperNOVA Film-slides, we achieved an order of magnitude greater sensitivity in a cytokine microarray assay than with … Continued