A Human-Machine Macromolecule Collaboration in Immunochemistry Laboratory, Contributing to Life-Saving Discoveries!
FARMOVS is a leading Clinical Research Organisation (CRO) with a unique advantage. With the only on-site ISO accredited and GLP certified Bioanalytical Laboratory on the African continent they offer the highest quality bioanalytical services for qualitative and quantitative drug, metabolite, and biomarker analysis in a variety of matrices. With advanced technology and backed by 30 years of bioanalytical experience, they have developed more than 550 validated analytical methods that adhere to ICH and FDA guidelines. Their analytical methods have been used in more than 3,000 pre-clinical and clinical trials, contributing to the manufacturing of pharmaceutical drugs that are now used by most households.
Their team of extraordinary Bioanalytical experts thrive on the excitement generated by new discoveries. Anton Engelbrecht, a subject matter expert in his field, manages the FARMOVS Immunochemistry Laboratory. Anton obtained his B. Med. Sc. Honours Degree in Analytical Pharmacology Research at The University of the Free State in 1999 and decided to invest his time in clinical research for the last 24 years. He explains that his daily journey in the world of clinical research does not merely consist of the analysis of macromolecules using various methods. It is a world filled with novel methods of analysis and subsequent technological integration that expands the horizons of clinical research. He feels that his work forms an important part of the discovery and production of new life-saving medicines that is constantly improving the quality of life of people all over the world.
Anton believes that new technology and innovation should be the building blocks of any laboratory. Thus, he recruited the best team members to join FARMOVS. Amongst these, are the three fastest members of our Immunochemistry Laboratory team, the STARLet Family! “We chose the Microlab® STARLet apparatus by Hamilton, because of its ability to perform sample analysis in large quantities at a greater speed by means of robotic pipetting and robotic automated microplate reading, which is a semi-automated process. The STARLet Family does the ‘heavy lifting’ for the rest of the team while we focus on enhancing the quality of the data as to ensure optimal results at all times. This effective collaboration adds substantially to the FARMOVS value offer”.
Below, Anton Engelbrecht and the Hamilton Microlab® STARLet.
“At FARMOVS we have the opportunity to work with the world’s top pharmaceutical companies where we can form part of the planning or manufacturing of labeled or conjugated antigens and antibodies specific for the drug of interest and then develop and validate a specific method. Alternatively, commercially available kits and reagents are also used for the same purpose if the Sponsor cannot supply the specific critical reagents. We work within a very well-planned and demanding schedule at the best of times.
“Luckily, our team can facilitate the processing of more than one thousand tubes daily. This is possible because our four Bioanalysts, two Biotechnicians, one Laboratory Assistant and the STARLet, WatsonTM and BioTek® families have fully optimized their workflow plan. Thereafter a team of six Data Bioanalysts perform independent data bioanalysis to ensure the integrity of the data. Planning and collaboration ensure that our work remains interesting and enjoyable, despite the high standards we maintain while running our laboratory at full capacity”, says Anton.
When asked what his daily life as a Bioanalysis Specialist entails, he was more than happy to share his knowledge. Anton beamed with enthusiasm regarding a recent improvement in the field of Immunoassay development for the purposes of pharmaceutical analysis. This involves the preparation of unique immunoanalytical reagents, analysis of new categories of compounds, methodology, and instrumentation. The most important examples in this field are the continuous development of Beat-based Immunoassays. This developing technology results in improvement of the analysis performance by increasing the sensitivity, decreasing the analysis time, simplification of the assay procedure, automation of the method, and miniaturisation of the analytical equipment.
Anton further shared that Immunoassay methods, such as Radioimmunoassay (RIA) and Enzyme Immunoassay (EIA) amongst others are also used at FARMOVS to analyse macromolecules for clients. “The RIA method is used for the determination of several pharmaceutically important compounds in biological fluids. An RIA requires a sample containing the antigen of interest, a complementary antibody, and a radiolabeled version of the antigen. To increase the selectivity of an assay, all samples are pre-treated to eliminate high molecular weight endogenous matrix components, including anti-drug antibodies”, explains Anton.
Anton indicated that a competitive RIA method is the most frequently used method by the Immunochemistry Laboratory. “A fixed concentration of labeled antigen, e.g. 125I-labeled antigen, also referred to as tracer and with a fixed volume of a pre-treated assay sample that contains the unlabeled antigen, are incubated with a constant volume of complementary antibodies in assay buffer. During incubation, the labeled and unlabeled antigens compete for the limited and constant antigen binding sites on the antibodies. The amount of tracer bound to the antibodies decreases as the concentration of the unlabeled antigen increases. The antibody-bound tracer is separated from the free tracer by precipitation.
“After decanting the free tracer, the antibody-bound activity of the sample (B) is measured by our other two impressive team members, the Wizard2 TM gamma counters and then calculated as a percentage (% B/B0) of the maximum binding (B0). Regression curve fitting is performed, and from this curve, the antigen concentration in unknown samples can be calculated by interpolation. All these calculations are performed using our famous WatsonTM Laboratory Information Management System (LIMS). A direct RIA method is also used by the Immunochemistry Laboratory if the presence of anti-drug antibodies must be evaluated”.
Anton explains that a fixed concentration of tracer is incubated with constant volumes of diluted samples. The tracer is typically the drug of interest that is 125I-labeled. During incubation, the tracer forms complexes with the anti-drug antibodies which may be present in the samples. The amount of tracer bound to the antibody increases as the concentration of the antibodies increases. This technique is perceived as ‘old-school’, but still stays the preferred method for very sensitive immunogenicity assays if 125I-labeled tracer is available”.
Anton further explained that the second method is an Enzyme Immunoassay (EIA), which is analogous to RIA except that the label is an enzyme rather than a radioisotope. “The basic approach for use of an enzyme as an Immunoassay label is appreciated by coupling an enzyme molecule to one of the immunoanalytical reagents (analyte or antibody), by a specific biochemical technique, and then carrying out the immunochemical reaction in the normal way. Following the separation of bound and free fractions of the analyte of interest, the remaining enzyme activity is monitored. This is achieved by adding substrate, and subsequent monitoring the turnover of the substrate to product.
“The product must possess measurable physical or chemical differences from the substrate. For example, colourless chromogenic substrates which are converted into coloured products by the action of the enzyme label. The coloured products can be simply measured by a Spectrophotometer. The measured signal is then correlated to the analyte concentration. Although an enzyme/chromogenic substrate system is frequently used, many other systems can also be used”.
Below, Anton Engelbrecht in the FARMOVS Immunochemistry Laboratory.
“To enable optimal analysis, we acquired a variety of the BioTek® Family of instruments. These include an Absorbance Reader and two models of Microplate Washers. The flagship, a Hybrid Reader from Biotek®, is also available for absorbance, luminescence, fluorescence, and time-resolved fluorescence detection. The Robotic Pipettors are also used by the Biotechnicians and Bioanalysts to dispense the reagents into the microplates.
Although we have adequate technology to provide market related results, we are planning to expand our team to include a Multiplex Platform that is a sensitive, fully automated Immunoassay platform with multiplexing and custom assay capability. This will pave the way to use an even more sensitive method to quantify biomarkers in the fields of oncology, neurology, cardiology, inflammation and infectious disease”. We aim to remain competitive in our industry, so naturally we must recruit the brightest and most evolved to join the team. Whether a human or a machine team member is irrelevant to us. Our clients deserve the best!”, Anton exclaimed.