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Home Uncategorized

Basics Of Using ELISA Kits For Bioprocess Optimization

by EmmaW
November 2, 2020
in Uncategorized
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Basics Of Using ELISA Kits For Bioprocess Optimization
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Whether it is an all-new biotherapeutic, chemical entity, or cellular/genetic therapy, a single drug’s development needs a significant amount of invested resources. Everything involved in the method, from its earliest discovery to delivery and production, should be wholly characterized, explored, and understood. That is because it makes it easier to prevent problems that result from cell death, cell stress, factors, and protein aggregation, affecting the reliable source of the drug’s manufacturer. Furthermore, ELISA kits are quite established for the process of contamination monitoring. It is essential to know that Elisa usually consumes a matched antibody pair for it to observe the availability of a particular analyte. Elisa happens to be completely sensitive and quantitative. However, the kits that are available in the market can detect through the picogram-per-mL levels.

Image source: https://www.pexels.com/photo/adult-biology-chemical-chemist-356040/

Even if someone works in upstream, drug discovery, or downstream bioprocessing, a definitive range of ELISA kits exists for the researchers. This happens to maintain the cell-line viability, monitor and optimize product integrity, and maximize the yield for a worthwhile final product.

Protein A

Protein A tends to be one of the bacterial proteins that are produced in Staphylococcus aureus. The protein A then enters the binding process where it binds to the IgGs along with affinity. Recombinant Protein A, however, has found an extensive utilization in the process of antibodies’ affinity purification and the commonly utilized visualization and immunodetection techniques. Such a process is majorly brought to use by specific industries in order to purify mAbs.

Moreover, the Protein A affinity chromatography is highly selective when it comes to mAbs, which, in turn, results in the purity that is more than than 95%, beginning from the complex samples. Such a technique went on to become an ideal method by obtaining significant yield and purity within one unit of operation. This further simplifies manufacturing operations and reduces the effort and time required to produce the purification process.

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Host-Cell Proteins

Host-Cell Proteins are the residual contaminant that usually appears in the biologic’s production. Organisms produce these types of cell proteins as their side product to build the intended protein. Since there are several possible host cells to choose from, many facilities want to use Chinese hamster ovaries or Escherichia coli (CHO) cells in the shape of model organisms.

Image source: https://www.pexels.com/photo/person-holding-petri-dish-3786217/

Even though the HCPs are essential for the functioning of natural cellular and some of them might be entirely harmless, at times, they are carried via such a purification process and considered to be undesirable to be present in the concluding product. Every year, a significant effort is put in through the industry to remove host-cell proteins from the biologic mixes. Such a process is further complicated by the availability of HCPs’ in meager quantities (commonly on the scale of ng/mL). This makes it more critical to quantify the residuals of HCP amid the biologics’ production. There are several techniques developed for the HCP analysis, like Western blot or gel electrophoresis. However, one of the most common methods is still ELISA. 

Final Thoughts

Whenever you pursue a protein’s production, the available introduction of the contaminants happens to be unavoidable. However, ELISAs tend to be equally suited for such an issue. ELISAs are specific and sensitive that enable manufacturing facilities to locate the availability of contaminants. Furthermore, the technology of ELISA is used to inquire about HCP and Protein A contaminants.

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