An attractive feature of using in vitro expression systems to generate biological compounds is the capability to produce a wide variety of compounds (proteins, peptides, antibodies) on various scales, in a cheap and rapid manner. Current methods to analyze any of these produced compounds require purification which poses a major drawback for working with products derived from expression systems. For example, a moderately abundant (soluble) protein can require 27 individual steps over four days to purify; a significant amount of time and expense to screen a single protein. Screening a library of potential biological compounds for therapeutic value, requires many purification steps for each compound before any analysis can be performed. This includes measurements as simple as determination of the mass of the produced biological compounds. To this end we have developed a unique DNA-based scheme using differential scanning calorimetry for determination of the masses of biological compounds in both purified and mixed media. Uniquely, our probes are universally applicable to a wide range of biological compounds enabling mass quantification prior to purification. This advance allows for initial analysis of biologics early in the production process, lowering associated time requirements and resource costs. An added benefits the possibility to supplant the numerous mass analysis kits currently on the market that rely on colorimetric or fluorescent methods. These methods are highly specific to the types of compounds assayed and have limitations preventing universal applicability. Our product offers a superior advantage of existing methods. In this project we plan to screen a library of biologically expressed SARS-CoV-2 spike and nucleocapsid proteins generated by our commercial collaborator. The aim of this work is to evaluate whether our assay is capable of predicting protein yields from their expression systems solely from a mass determination of their unpurified samples. Additionally, we will use our assay to verify the protein structure before and after purification.
Public Health Relevance Statement: Project narrative: Production of many biopharmaceuticals or biologics, low abundance and engineered molecules is accomplished using prokaryotic or eukaryotic expression systems; allowing for the manufacture of a large variety of biological compounds on a small scale for screening applications, and large scale on the metric-ton scale for therapeutics. However, while expression of the targeted biological compounds is cheap and fast, a major drawback is the time required and expense involved in purification and isolation of targeted compounds. We have developed an assay that will allow quantitative analysis of proteins prior to purification; in this project we will screen biologically expressed SARS-CoV-2 proteins from a commercial collaborator for validation studies.
Project Terms: Antibodies, Biological Assay, Assay, Bioassay, Biologic Assays, Biological Products, Biologic Products, Biological Agent, biologics, biopharmaceutical, biotherapeutic agent, Buffers, Differential Scanning Calorimetry, Calorimetric Differential Thermal Analysis, Clinical Trials, Culture Media, growth media, DNA, Deoxyribonucleic Acid, Pharmaceutical Preparations, Drugs, Medication, Pharmaceutic Preparations, drug/agent, Engineering, Environment, Equipment, Feedback, Fluorescence, Glass, In Vitro, Laboratories, Libraries, Methods, Painful, Pain, Peptides, Production, Proteins, diagnostic kit, test kit, Diagnostic Reagent Kits, Reagent, Research Resources, Resources, Mass Photometry/Spectrum Analysis, Mass Spectrometry, Mass Spectroscopy, Mass Spectrum, Mass Spectrum Analyses, Mass Spectrum Analysis, Survey Instrument, Surveys, Testing, Thermodynamic, Thermodynamics, Time, Viral Proteins, Viral Gene Products, Viral Gene Proteins, virus protein, Work, base, Vial device, Vial, Phase, Biological, biologic, Individual, Measurement, Collaborations, Therapeutic, instrument, Complex, Scanning, Techniques, System, Nucleocapsid Proteins, experience, melting, novel, validation studies, Sampling, Property, response, Complex Mixtures, preventing, prevent, Address, protein structures, proteins structure, protein structure, Molecular Target, Protein Analysis, Scheme, Characteristics, Process, Modification, Instruction, cost, design, designing, Outcome, Consumption, Peptide antibodies, novel therapeutics, new drug treatments, new drugs, new therapeutics, new therapy, next generation therapeutics, novel drug treatments, novel drugs, novel therapy, data reduction, screening, 2019-nCoV, 2019 novel corona virus, 2019 novel coronavirus, COVID-19 virus, COVID19 virus, CoV-2, CoV2, SARS corona virus 2, SARS-CO-V2, SARS-COVID-2, SARS-CoV-2, SARS-CoV2, SARS-associated corona virus 2, SARS-associated coronavirus 2, SARS-coronavirus-2, SARS-related corona virus 2, SARS-related coronavirus 2, SARSCoV2, Severe Acute Respiratory Coronavirus 2, Severe Acute Respiratory Distress Syndrome CoV 2, Severe Acute Respiratory Distress Syndrome Corona Virus 2, Severe Acute Respiratory Distress Syndrome Coronavirus 2, Severe Acute Respiratory Syndrome CoV 2, Severe Acute Respiratory Syndrome-associated coronavirus 2, Severe Acute Respiratory Syndrome-related coronavirus 2, Severe acute respiratory syndrome associated corona virus 2, Severe acute respiratory syndrome corona virus 2, Severe acute respiratory syndrome coronavirus 2, Severe acute respiratory syndrome related corona virus 2, Wuhan coronavirus, coronavirus disease 2019 virus, coronavirus disease-19 virus, hCoV19, nCoV2, ACE2, angiotensin converting enzyme 2, angiotensin converting enzyme II
