Antibody purification is a multistep process by which antibodies with high purity can be achieved. Antibodies are widely used as injectables and parenteral products for human use. Monoclonal antibodies dominate the antibody field.
2. What is Antibody Purification??
• Antibody purification is a multistep process by which
antibodies with high purity can be achieved.
• Antibodies are widely used as injectables and parenteral
products for human use.
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5. Why to Purify Antibody?
• To remove possible contaminants serum proteins such as
albumin, transferrins and cell degradation products like DNA
and cellular proteins
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6. Purification Methods
The choice of a purification method is based on a these factors:
• Nature of antibody,
• Nature of feedstock,
• Scale of production,
• Economics – cost and other factors,
• Process Timings, and
• Desired purity.
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8. Antibody Purification:
Step 1: Sample Preparation
• It is the initial step in which crude protein sample is
conditioned or making it ready for the initial capture step.
• Involves changing pH or Ionic strength, dilution of the crude
sample or addition of salts for the ionic strength. (Cost
effective???)
• Buffer exchange by size exclusion chromatography or to use
ultrafiltration or diafiltration.
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10. Antibody Purification:
Step 3: Secondary Purification
• On the nature and the optimization requirement of the crude
antibody source.
• In addition to protein contaminants, other impurities such as
DNA, endotoxins, viruses, and aggregates need to be
removed. In such cases, a multistep procedure is almost
inevitable
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11. Antibody Purification:
Step 4: Polishing / Formulation
• Final polishing / formulation step can be considered as a part
of purification in which it removes conditions that would
impair the stability or utility of the antibody in its intended
use.
• Ultrafiltration.
• SEC , Diafiltration.
• Lyophilization.
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