We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress hp
Sign In
Advertise with Us
PURITAN MEDICAL

Download Mobile App




Peptides for Immunoenzymatic Assays Immobilized on Microplates

By LabMedica International staff writers
Posted on 28 Aug 2012
Synthetic peptides are widely used in indirect enzyme-linked immunosorbent assays (ELISA) to detect and characterize specific antibodies in clinical samples. More...


A simple, fast, and inexpensive protocol has been described that shows how immobilized synthetic peptides can be bound to plastic surfaces for standard ELISA, which could be used as an alternative to the common technique of conjugating peptide antigens to carrier proteins.

Scientists at the University of Sassari (Italy) devised the technique, which is based on the use of maleimide-activated bovine serum albumin or keyhole limpet hemocyanin as a protein anchor adsorbed on the polystyrene surface of the microtiter plate. Following adsorption of the carrier protein, sulfhydryl-containing peptides are cross-linked with an in-well reaction, allowing their correct orientation and availability to antibody binding, avoiding the time consuming steps needed to purify the hapten–carrier complexes.

Conventional in-solution maleimide-mediated coupling reaction (ISC) was performed by using the Imject Maleimide Activated Immunogen Conjugation kit (Pierce; Rockford, IL, USA). Nine synthetic peptides (Nurex; Sassari, Italy) with length ranging from 10 to 15 amino acids were designed to display different hydrophobic/hydrophilic characteristics and to carry either an N or C terminal cysteine, to allow cross-linking with maleimide-activated carrier proteins. The immunoreactivity of peptides was tested by using both monoclonal and polyclonal antibodies in standard ELISA assays, and compared with established coating methods.

The efficiency of direct cross-linking method (DCLM) was tested with four different peptides and subsequent standard indirect ELISA with polyclonal antibodies was performed. All the sera specifically recognized the corresponding specific antigens when directly cross-linked onto the multiwell plate. Results in terms of signal strength were comparable to that obtained with ELISAs performed by using plates coated with ISC conjugates.

The authors concluded that DCLM proved to be simple, reproducible, cost-effective, and suitable with peptides of different lengths and hydrophobic/hydrophilic characteristics. The DCLM can be considered a straightforward, faster, and much more convenient alternative to standard coupling protocols, allowing saving time and cut costs in ELISA applications. When considering its time- and cost-effectiveness, DCLM may prove to be applicable also to automated systems for high-throughput peptide screenings. The study was published in the August 2012 edition of the Journal of Immunological Methods.

Related Links:

The University of Sassari
Pierce
Nurex



Gold Member
Flocked Fiber Swabs
Puritan® Patented HydraFlock®
Serological Pipet Controller
PIPETBOY GENIUS
New
PSA Assay
CanAg PSA EIA
New
Staining System
RAL DIFF-QUIK
Read the full article by registering today, it's FREE! It's Free!
Register now for FREE to LabMedica.com and get access to news and events that shape the world of Clinical Laboratory Medicine.
  • Free digital version edition of LabMedica International sent by email on regular basis
  • Free print version of LabMedica International magazine (available only outside USA and Canada).
  • Free and unlimited access to back issues of LabMedica International in digital format
  • Free LabMedica International Newsletter sent every week containing the latest news
  • Free breaking news sent via email
  • Free access to Events Calendar
  • Free access to LinkXpress new product services
  • REGISTRATION IS FREE AND EASY!
Click here to Register








DIASOURCE (A Biovendor Company)

Channels

Hematology

view channel
Image: CitoCBC is the world first cartridge-based CBC to be granted CLIA Waived status by FDA (Photo courtesy of CytoChip)

Disposable Cartridge-Based Test Delivers Rapid and Accurate CBC Results

Complete Blood Count (CBC) is one of the most commonly ordered lab tests, crucial for diagnosing diseases, monitoring therapies, and conducting routine health screenings. However, more than 90% of physician... Read more

Pathology

view channel
Image: A real-time trial has shown that AI could speed cancer care (Photo courtesy of Campanella, et al., Nature Medicine)

AI Accurately Predicts Genetic Mutations from Routine Pathology Slides for Faster Cancer Care

Current cancer treatment decisions are often guided by genetic testing, which can be expensive, time-consuming, and not always available at leading hospitals. For patients with lung adenocarcinoma, a critical... Read more

Technology

view channel
Image: Researchers Dr. Lee Eun Sook and Dr. Lee Jinhyung examine the imprinting equipment used for nanodisk synthesis (Photo courtesy of KRISS)

Multifunctional Nanomaterial Simultaneously Performs Cancer Diagnosis, Treatment, and Immune Activation

Cancer treatments, including surgery, radiation therapy, and chemotherapy, have significant limitations. These treatments not only target cancerous areas but also damage healthy tissues, causing side effects... Read more
Copyright © 2000-2025 Globetech Media. All rights reserved.