deltaDOT Techniques

 Monoclonal Antibodies Profiling

In order to assess the bioactivity and safety of recombinant monoclonal antibodies (mAbs) as biological drugs, it is important to fully characterise them. Not only is this information crucial for the assessment of the suitability of a potential biopharmaceutical product, it is also important for the optimisation and QA/QC of the manufacturing process, storage conditions and life-time assessment of the drug.

HPCE comparison to MS

Ribonuclease B glycoform separations were used as benchmark molecule in a comparative study of analytical techniques used in glycosylation profiling. Techniques compared included deltaDOT’s HPCE system and Mass Spectrometers. The data presented here compares deltaDOT’s HPCE with the Agilent 6530 Q-TOF system.

Antibiotic Analysis

Analysis of ß-lactam antibiotics in food products can be performed using the deltaDOT HPCE. This enables the possibility of on-site analytics of at food at source or at distribution centres. Various penicillin derivatives are widely used to treat infectious diseases in humans and animals, acting to kill or inhibit the growth of the infectious organism. A mixture of these 8 antibiotic derivatives was analysed using Micellar Electrokinetic Capillary Chromatography.

 Nicotine analysis

DARPA started Project Blue Angel, part of which was the Accelerated Manufacture of Pharmaceuticals (AMP) program. Following two scoping phases, the Tobacco plant Nicotiana benthamiana was selected as the expression system of choice for the emergency product of vaccines following a pandemic such as predicted in the concurrent Swine Flu crisis. While this predicted pandemic did not materialise, several facilities where constructed for tobacco based pharmaceutical production and some have been retasked to produce vaccines and future personalised medicine products. The use of plants for drug production allows improved yield per dollar over other systems, but also presents demanding purification issues.

 Glatiramer Acetate Analysis

Glatiramer acetate is an immunomodulator treatment for multiple sclerosis. It is a blend of peptides that are composed of the four amino acids glutamic acid, lysine, alanine, and tyrosine that comprise the Myelin basic protein. This is the protein that initiates the autoimmune Multiple Scelerosis event when its liberation from its normal lipid rich environment causes conformational changes. The peptides are generated by a Trypsin digest and then need accurate analysis to ensure a random distribution across the range of possible peptides in the formulation. The random-sized peptides appear to sequester the antibodies from the autoimmune response, acting as a decoy molecule and initiating the induction of T regulatory cells away from the myelin sheath.

 Amino Acid Analysis

Amino acids are the building blocks of proteins. There are 20 standard amino acids that are genetically encoded to form the proteins that comprise and are in control of the cell. In biopharmaceuticals proteins are built in expression systems such as mammalian cells. This done by the incorporation of amino acids that have been coded for genetically engineered DNA, mediated by RNA. This key to the production of biopharmaceuticals and the analysis of amino acids is vital in different stages on the bioprocess. There is a large and growing market for these drugs and their efficient production is crucial.

 Protein Fingerprinting

Protein profiling of biologics can be time-consuming, expensive, inconclusive and subjective. The profiling or fingerprinting of complex analytes with multiple proteins present is increasingly important, however, in the biopharmaceutical sector. These applications are not confined to the production of therapeutic molecules such as monoclonal antibodies and other protein products; In the discovery phase of emerging and established therapies, such as stem cell therapy production, viral gene vector characterisation and synthetic biology systems optimisation, the need for higher quality data is also vital.

Water analysis

Water is a key component in biopharmaceutical production. It is used in synthesis, purification, reconstitution, fill and finish and finally in waste removal and the cleaning of all vessels and piping in non-single use and hydroponic plant facilities. Different grades of water may be used for different processes and quality control at each of these units is vital and expensive. Control of the ionic content, conductivity and microbacterialogical loads are all important parameters and while the latter two are established the former is not, even though data exists to suggest that ionic content is important in e.g. final product stability.