R&D

Why Proteomics?

Proteome is a word made up of words “protein” and “ome”, and means a set of proteins in a sample (tissue, organ, cell, body fluid, etc.) at a specific time and specific condition.

2-D electrophoresis system that effectively analyzes all the proteins in a sample with high reproducibility.
Development of mass analyzing method that allows a small amount of protein from 2-DE gel to be identified in a short time. Development of bioinformatics allows quick comparison between the information in the given protein sample and that in the on-line data.

Genetic sequence was revealed but many biological unexplainable condition is still existing.
It is able to explain from protein research that is a final substance that Genetic is a lot of conducting.
Genetic diseases only includes 2~3% of all diseases, so there is certain limitation to development of diagnosis and treatment of diseases using genomics.
But protein is a final substance indicating biological condition and adjusting in contrast to DNA and RNA.
Sometimes, PTM product is more important than pure protein.
It is able to explain a disease from protein and modification of proteins.

The above tells us that proteomics technology, compared to genomics technology, can create high value in a short time.

-open range in purpose of interest
-ability to directly observe complicated phase change in a balanced comparison
-ability to isolate and identify a very small amount of protein (<10fmol) from cells or complicated mixture of proteins.

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Mass Spectrometry

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Mass spectrometry (MS) is an analytical technique that measures the molecular weight of molecules based upon the motion of a charged particle in an electric or magnetic field. It is used for determining masses of particles, for determining the elemental composition of a sample or molecule, and for elucidating the chemical structures of molecules, such as peptides and other chemical compounds. This technique is performed by a mass spectrometer. The MS principle consists of ionizing chemical compounds to generate charged molecules or molecule fragments and measuring their mass-to-charge ratios (m/z).

In a typical MS procedure
1. A sample is loaded onto the MS instrument, and undergoes vaporization.
2. The components of the sample are ionized by one of a variety of methods (e.g., by impacting them with an electron beam), which results in the formation of charged particles (ions)
3. The ions are separated according to their mass-to-charge ratio in an analyzer by electromagnetic fields..
4. The ions are detected, usually by a quantitative method.
5. The ion signal is processed into mass spectra.

Generally MS instruments consist of three modules
ㅁ An ion source, which can convert sample molecules into ions.
(or, in the case of electrospray ionization, move ions that exist in solution into the gas phase)

ㅁ A mass analyzer, which sorts the ions by their masses by applying electromagnetic fields.
ㅁ A detector, which measures the value of an indicator quantity and thus provides data for calculating the abundances of each ion present.

The technique has both qualitative and quantitative uses. These include identifying unknown compounds, determining the isotopic composition of elements in a molecule, and determining the structure of a compound by observing its fragmentation. In the field of protein identification and proteomics, a mass spectrometer is judged upon sensitivity, mass accuracy and tandem mass spectrometry.

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Immuno Assay

Enzyme-Linked ImmunoSorbent Assay (ELISA) is frequently used for measuring of a particular protein in sample. There are two main variations on this method: it can be determine how much antibody is in a sample, or it can be determine how much protein is bound by an antibody.

Direct Enzyme-Linked ImmunoSorbent Assay (ELISA)

The direct ELISA is a method for detecting and measuring antigen concentration in a sample. A particular antigen in a sample is detected directly by a capture antibody. Microplates are coated with a sample containing the target antigen, and the binding of labeled antibody is quantitated by a colorimetric, chemiluminescent, or fluorescent end-point.
It is a relatively quick method because the secondary antibody is not involved. However, the direct ELISA requires the labeling of every antibody to be used, which may be a time-consuming and additional cost.

However, there are a few antibodies that may be unsuitable for direct labeling and no additional signal amplification that can be achieved by secondary antibody in many assay methods.

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Indirect Enzyme-Linked ImmunoSorbent Assay (ELISA)

The indirect ELISA uses a labeled secondary antibody for detection.
After a primary antibody is incubated with the antigen, a labeled secondary antibody that recognizes the primary antibody binds is added to
microplates. The antigen is then quantitated by measuring the amount of labeled second antibody bound to the matrix, through the use of a colorimetric, chemiluminescent, or fluorescent substrate. It is also useful to measuring the concentration of antibody in sample
(serum, plasma, blood etc.)

Sensitivity is increased because each primary antibody contains several epitopes that can be bound by the labeled secondary antibody, allowing
for signal amplification. A commercially available secondary antibody can
be used and immunoreactivity of the primary antibody is not affected by labeling.

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Sandwich ELISA

The sandwich ELISA measures the amount of antigen between two layers of antibodies. It is restricted to the quantitation of multivalent antigens because two different antibodies involved in binding to antigen. After the “capture” antibody is bound to a solid phase typically attached to the
bottom of a plate well, antigen is then added and allowed to complex with the bound antibody. Unbound materials are then washed away, and a labeled “detection” antibody is allowed to bind to the antigen,thus completing the “sandwich”.

Both the capture and the detection antibody must recognize separate epitopes on the antigen so that they do not hinder each other’s binding.
The assay is then quantitated by measuring the amount of labeled second antibody bound through the use of a colorimetric, chemiluminescent, or fluorescent substrate. Sandwich ELISAs for quantitation of antigens are especially valuable when the concentration of antigens is low and/or they are contained in high concentrations of contaminating protein.

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Competitive ELISA

Competitive ELISA is a competition between two antigens or two antibodies to the same binding site in an antigen-antibody reaction. The design of the diagnostic method depends on the substance to be measured. As the quantity of Inhibitor Antigen, which has the same reactive site for a specific antibody as in the schematic diagram, increases, the signal detected becomes lower.

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Rapid Assay

Lateral flow immunochromatographic assay (LFIA) is used to detect the presence of trace amounts of analyte in various samples (serum, plasma, urine, swear, and other biofluids) using antigen – antibody reaction(immune reaction).  Rapid test diagnostic kits that are manufactured using LFIA technology is widely used in the field of point of care test(POCT) to analyze qualitatively and quantitatively the analyte in a short time by one-step experiment. Typical example, there is a widely known home pregnancy diagnostic kit for general consumers, and not only  that is used  to the diagnosis and examination of various diseases such as diagnosis of viral infection, medicine, Agriculture, livestock industry, food, military, and the environment.

The Rapid test kit, which is produced using the Lateral Flow immunochromatographic assay (LFIA), can be used an analyte-specific binding antibody immobilized on the membrane and another analyte-specific binding antibody with ligand capable of color development to detect the analyte. The ligand colorimetric intensity according to the concentration of the analyte on the membrane can be measured by a simple POCT device or by visual inspection for getting the qualitatively  or quantitative analysis results of analyte. Rapid kit strips are made by assembling sample pads that absorb the sample, a ligand-antibody conjugate pad, an antibody-immobilized membrane, and a absorbsion pad that absorbs the remaining sample.

LFIA Rapid kit scheme

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The analyte in the sample reacted with the ligand(Gold nano particle)-antibody conjugate was determined to be positive by color development with a specific antibody fixed at the position of the test line of the membrane (Sandwich immunoassay). If there is no analyte present in the sample, the line will not appear on the test line and will be judged as negative. In addition to the control line, other antibodies are immobilized on the membrane which indicates the normal termination of the reaction, regardless of the presence or absence of analytes in the sample.

Test Results/ Positive

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Proteometech is to overcome the hook effect (prozone effect), developed an innovative and new quantitative rapid test assay which has the competition assay methods (alternative detection line) and sandwich immunoassay methods(classical test line) in a test. Our assay method showed that we can detect more broad range of target molecules than classical rapid detection method without a hook effect. This new technology is an easy and convenient method which is suitable to measure the target with a wide dynamic range. We have developed and commercialized an ImmunCheck ™ IgG kit and a pregnancy diagnosis kits. ImmunCheck ™ IgG kit can quantitatively measure the concentration of IgG in the serum, pasma, whole blood by using the principle, and are developing additional products.

Classical rapid kit reaction

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Reaction scheme of LFIA Rapid kit using new technology

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Reaction scheme of LFIA Rapid kit using new technology

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Biomarker Discovery

A biomarker, or biological marker,is in general a substance used as an indicator of a biological state.
It is a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention. Biomarker discovery has been accelerated by proteomics, which study to entire protein expression at any given time in cell or tissue. If a certain protein is the functional molecule in cells, measurement of the differential expression of proteins could indicate disease-specific changes in tissues or organs.

Candidate biomarkers identified by proteomic profiling of serum and tissue could be used for diagnosis and/or prognosis of disease. We have tried to discover novel biomarkers which are related to several kinds of cancer and infectious disease.

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Biomolecular Imaging

Biomolecular imaging enables the visualization of the cellular function and the follow-up of the molecular process in living organisms without perturbing them. The multiple and numerous potentialities of this field are applicable to the diagnosis of diseases such as cancer, and neurological and cardiovascular diseases. Radioactive materials such as F-18, Ga-67, Tc-99m, I-123, I-131 and Ti-201 are frequently used to diagnosis and/or prognosis of disease. Many fluorescent dyes have also been studied for molecular imaging but these dyes might cause safety issues on human.

We are developing a natural dye which is conjugated with bioprobes. Since our dye is from natural biomolecule,
it may not induce toxic effects on human.

Biosensor

What is biosensor? An analytical device for the detection of an analyte that combines a biological component with a physicochemical detector component

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Principle of biosensor

– Biosensors generally consist of a bioreceptor that reacts with the analyte, a signal transducer and an output that shows the measurement signal.
– Detectors, called bioreceptor, use substances that mimic organisms or biological elements to interact with analytes.
– The interaction between the analyte and the bioreceptor takes place in a signal transducer capable of measuring the response signal.

Core technology

Inspection Chip

  • Optimization of aptamer coating
  • Coating uniformity of probe materials

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전용 검사 시스템 개발

  • High quality of inspection chip
  • Detection of bacterial and antibiotic susceptibility tests using clinical samples

Development of real-time antibiotics
susceptibility inspection system

Application field

Pharmaceutical field

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Antibiotic screening and antibiotic drug development application

Medical field

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Effective prescription of antibiotic

Animal antibiotic field

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Livestock and companion animals Antibiotic susceptibility test

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