ABSTRACT
With the existence of several thousand unique human allergens, a multiplex format, such as protein microarrays, is an attractive option for allergy screening. To determine the feasibility and sensitivity of using an enzyme-based, colorimetric protein microarray assay, three common allergens (mold, dust-mite, grass) were arrayed and added sera assayed for responsive human IgE. Normal, low positive, and negative control samples were assayed to determine optimal reaction parameters. Sensitivity of the assay (in international units, IU) was determined by constructing a standard curve using World Health Organization (WHO) standards. The system described here can reliably detect allergen-specific IgE below 0.35 IU, the current WHO standard cutoff. By taking advantage of the sensitivity of enzyme-linked immunosorbent assays (ELISAs) and the multiplex format of microarrays, we have achieved a high throughput system, capable of screening patients for allergen-susceptibility with optimal sensitivity.
Subject(s)
Allergens/administration & dosage , Immunoglobulin E/analysis , Protein Array Analysis/methods , Animals , Case-Control Studies , Colorimetry , Dust/immunology , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/statistics & numerical data , Fungi/immunology , Humans , Hypersensitivity, Immediate/diagnosis , Hypersensitivity, Immediate/immunology , Immunoglobulin E/blood , In Vitro Techniques , Mites/immunology , Phleum/immunology , Protein Array Analysis/standards , Protein Array Analysis/statistics & numerical data , Rhinitis, Allergic, Seasonal/diagnosis , Rhinitis, Allergic, Seasonal/immunology , Sensitivity and SpecificityABSTRACT
The advent of pathotropic (disease-seeking) targeting has transported genetic medicine across the threshold of history with the progressive clinical validation of Rexin-G, a tumor-targeted nanosized anti-cancer agent. Achieving noteworthy single-agent efficacy and survival benefits in otherwise intractable cancers, the molecular biotechnology platform has stimulated intense interest in the underlying mechanisms-of-action. This report exhibits the effective localization of Rexin-G nanoparticles within a metastatic liver lesion, as observed upon its surgical excision.
Subject(s)
Adenocarcinoma/drug therapy , Antineoplastic Agents/therapeutic use , Cyclin G1/genetics , Genetic Therapy/methods , Liver Neoplasms/drug therapy , Pancreatic Neoplasms/drug therapy , Adenocarcinoma/secondary , Clinical Trials, Phase I as Topic , Clinical Trials, Phase II as Topic , Genetic Vectors , Humans , Liver Neoplasms/secondary , Nanoparticles/therapeutic use , Pancreatic Neoplasms/pathology , Plasmids , RetroviridaeABSTRACT
We have developed an ISA (individual-specific autoantibody) identification method to identify biological samples based on an individual's unique class of autoantibodies. This method involved the presentation of human proteins derived from crude lysates after SDS/PAGE separation and transferance to a solid support. In the present study, ISA strips are produced and developed on a new protein microarray. In making the ISA strips, it was found that variation in protein migration during electrophoresis and strip-manufacturing quality control limit the reliability of the assay. Therefore it was decided to semi-purify and separate proteins by column chromatography in large batches and to develop an ISA-specific protein microarray. It was found that this ISA protein microarray approximates a similar serum titre as the ISA strip and is predicted to circumvent the batch-to-batch production issues related to SDS/PAGE.