Antibody Purification Technology

Antibody purification is the process of extracting antibodies from an antiserum (pAb), ascites fluid or cell culture supernatant of a hybridoma cell line (mAb). The purification can be performed without affinity tag by the physicochemical properties, antigen-specific affinity or the antibody class. Physicochemical properties are the molecular weight, charge, or clusters of specific residues. According to the origin of the sample, purification with the help of physicochemical properties or antigen-specific affinity can cause the isolation of further proteins and antibodies with similar properties besides the target antibody.

Fig. 1 The basic flowchart for antibody purification.

• Affinity Chromatography
  Affinity Chromatography is a biochemical separation and purification technique with reversible reactions, such as antigens and antibodies. It is suitable for purifying the target substance from the mixture with complex composition and impurity content much larger than the target substance.
• Ion Exchange Chromatography
  Ion exchange chromatography is one of the methods for separating macromolecules with similar properties from complex mixtures. Since proteins have isoelectric points, when proteins are in different pH conditions, their charged states are also different.
• Size Exclusion Chromatography
  Size exclusion chromatography is also known as size exclusion chromatography (SEC). It is mainly based on the relative relationship between the pore size of the gel pores and the coil size of the polymer sample molecules to separate the solute. It is mainly used to separate compounds with large molecular weight differences.
• Hydrophobic Interaction Chromatography
  Hydrophobic interaction chromatography is a chromatographic method for separation using the difference in hydrophobic force between the hydrophobic groups of sample molecules and the hydrophobic ligands of the chromatography medium in the salt-water system.

• Beneficial to preserve and exclude the influence of miscellaneous proteins on the results.

• Provide a consistent and inexpensive source of reagents for a variety of biomedical research applications.
• Treat a variety of diseases including cancer, autoimmune, metabolic and infectious diseases, using antibody drugs.

Laboratory Centrifuges

A centrifuge is any device that applies a sustained centrifugal force—that is, a force due to rotation. The widest use of centrifuges is for the concentration and purification of materials in suspension or dissolved in fluids. Suspended particles denser than the suspending liquid tend to migrate toward the periphery, while those less dense move toward the centre.

Chromatography Columns

Columns play a central role in the performance of chromatographic process. Well-packed columns with consistent performance in the whole design space of a process will deliver consistent product recovery and separation from contaminants. Proper design of columns delivers increased resolution between peaks facilitates the packing process of multiple resin types at different bed heights, and provides scalability from process development to full-scale manufacturing.

Spectrophotometers

Spectrophotometer is an optical instrument that measures the intensity of light in relation to the wavelength. It can measure just about anything such as: plastics, paper, liquid, metal, fabric and so on. A spectrophotometer is used to check for the quality of product in a production facility and identify and quantify microscopic samples in analytical laboratories.

pH Meters and Osmometers

A pH meter is a scientific instrument that measures the hydrogen-ion activity in water-based solutions, indicating its acidity or alkalinity expressed as pH. The pH meter measures the difference in electrical potential between a pH electrode and a reference electrode. The pH meter is used in many applications ranging from laboratory experimentation to quality control.

Chromatography Fraction Collector

The fraction collector is typically a rotating rack that can be filled with test tubes or similar containers. It is an essential component for liquid chromatographs for the purpose of separation in preparative chromatography. It allows samples to be collected in fixed volumes, or can be controlled to direct specific fractions detected as peaks of protein concentration, into separate containers.

Chromatography Refrigerator

The chromatography refrigerator is a device that is used to freeze, cool, and hold samples of liquids that are being studied in research, clinical, and industrial laboratories. The studied liquid may be anything from a rare earth metal to a plant extract. The purpose of the refrigerator is to ensure that the temperatures of the sample stay at a constant low temperature without melting or boiling.

Electrophoresis Systems

Electrophoresis equipment applies an electric charge to molecules, causing them to migrate towards their oppositely charged electrode. The technique is found in all research and clinical laboratories utilizing DNA and protein applications, and is divided into gel and capillary techniques. Choice of electrophoresis equipment depends on the molecule of study, appropriate method of separation and downstream application.

Chromatography Systems

Chromatography systems include all the equipment needed for sample separation, a power source and program software. Important considerations when selecting a chromatography system are the chemical and physical properties of the substance such as chemical reactivity, size, mass and shape and requirements of the laboratory, such as flow process, validation requirements and sample tracking.

STEMart provides you with a variety of antibody preparation and purification technology equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for antibody purification technology, please feel free to contact us.