Antibody generation raises a number of questions… what is the difference between their classes, forms and types? And how do recombinant monoclonal antibodies (rAbs) overcome the drawbacks of classical monoclonal antibodies (mAbs)?

Monoclonal antibodies are ubiquitous in biomedical research and medicine. They are used to fight, diagnose and research disease and to develop and test new drugs. The antibodies are divided in 5 classes or isotypes, several subtypes and forms and can be generated in vivo or in vitro.

In this short article, my aim is to define what an antibody is, to highlight the differences between a hybridoma monoclonal antibody and a recombinant monoclonal antibody and to point out how rAbs bring solutions to the classical drawbacks of mAbs.

Biologicals are increasingly being used in therapy, helping to cure thousands of patients, or at least improving their quality of life.

Research continues to move forward in this area, as proved by 7 400 citations in 2014 only when using “biologicals” as a search term, or about 200 citations when using “therapeutic mAb”. Many research groups, especially those working on Oncology, continue to investigate on the mechanism of action of biologicals, specially mAbs, as well as on any possible resistances found in non-responsive patients.

For these mechanistic or functional studies, antibody-based techniques such as ELISAs, antibody arrays, flow cytometry, etc. are routinely performed, using samples from patients being treated with therapeutic mAbs.

Glycation of proteins in blood circulation occurs naturally when circulating sugar is metabolized in the body. Glucose is known to attach to hemoglobin in the bloodstream to form glycated hemoglobin (HbA1c).

Glucose and Glycated hemoglobin (Bb1Ac) as diabetic biomarkers

Hemoglobin is a long-living blood protein complex (around 120 days). Such stability can be used to measure blood HbA1c and thus, indirectly monitor average blood glucose levels during a longer period of time than daily glycemia measurements.

A possible allegory would be to compare Hb1Ac measurements with a movie covering a period of several weeks (average hyperglycemia),  and glucose blood testing with instant photography (punctual glycemia with possible fluctuation over the day).

Glycemia and Hb1Ac measurements are complementary data. They can help diagnose, monitor and stratify patients with diabetes (eg. American Diabetes Association, World Health Organization…). It might also help in the design of personalized therapies. Diabetes.co.uk mentions 2 clinical studies which show that improving HbA1c by 1%, might reduce risks of microvascular complications by 25% for patients with type 1 diabetes or type 2 diabetes.

Direct ELISA with anti-HbA1c clone 75C9

Immunoassays are robust antibody-based quantitative methods. They are used to measure glycated hemoglobin HbA1c concentration expressed as a percentage of HbA1c to total hemoglobin.

These immuno-assays require well characterized and reliable primary antibodies.

A new hybridoma clone (75C9) specific for HbA1c has been released.

This 75C9 clone has been validated to measure Hb1Ac as a diabetic biomarker by direct and sandwich ELISA and complements the clone Hb6 which reacts with both unmodified and glycated hemoglobin.

Want to develop hemoglobin-specific ummunoassays with reliable monoclonals?

HbA1c calibration curve with Hb6 and 75C9 clones in fluoro-immunoassay.

This is something the experts at tebu-bio can help with, you might like to get in touch with them for solutions regarding your research projects.