Isolation and Purification of Bioactive Proteins from Bovine Colostrum

Wu, Mianbin; Wang, Xuewan; Zhang, Zhengyu; Wang, Rutao

doi:10.5772/20246

Cited by 3 publications

(3 citation statements)

References 40 publications

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“…27 Accordingly, selective elution of ion-exchange resins is achieved by slowly increasing the ionic strength (to competitively disrupt ionic interactions between the column matrix and the proteins) or by pH change (to lose the charge of reactive groups on the protein surfaces). 42 In this regard, this method has also been used for virus purification in our study.…”

Section: Antigen Fractionationmentioning

confidence: 94%

“…11 Precipitation method is an effective approach for protein concentration based on their sensitivity to the ionic strength, different pI, and other features. 42 For example, Liu et al 22 purified the produced anti-HAb18G/CD147 pAbs in rabbits using the ammonium sulfate precipitation and antigen-immunoaffinity chromatography methods. These purified antibodies showed a good performance in sandwich ELISA.…”

Section: Sds-page Of the Produced Antibodymentioning

confidence: 99%

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Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat

et al. 2022

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Purpose: Among all known human coronaviruses, some viruses (e.g., SARS-CoV-2) cause severe pneumonia or even death. With the regard to its spread and the importance of its rapid identification/treatment, and because pAbs are relatively cheap, able to bind to more sites on antigens and even neutralize them, this study was done for the production and purification of anti-SARS-CoV-2 polyclonal antibodies (pAb) in rats. Methods: Viral antigen purification was performed by PEG/NaCl precipitation. The efficiency of the sucrose cushion method was also investigated to produce a purer antigen. Immunization was done and antibody purification was performed by ammonium sulfate precipitation (33%), dialysis, and ion-exchange chromatography. Western blotting and ELISA were performed to verify the antibody specificity. All data were analyzed by SPSS software. Results: The results showed that the amount of concentrated virus increased with the increase of PEG concentration. Moreover, the sucrose cushion method increased its purity. Besides, induction of immune response in rats for pAb production with high titers was reached via these antigens and ELISA/western blot results indicated a suitable antibody-antigen interaction. Additionally, it was shown that ion-exchange chromatography could be a suitable technique for IgG purification. Conclusion: Herein, we presented a simple and cheap method for the purification of whole viral particles with relatively high quality. The results verified that these antigens could elicit a good immune response in the rat. The obtained pAbs showed a good specificity toward SARS-CoV-2 antigens. Accordingly, this study proposes a promising method for viral vaccine development.

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Section: Antigen Fractionationmentioning

confidence: 94%

Section: Sds-page Of the Produced Antibodymentioning

confidence: 99%

Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat

et al. 2022

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“…The binding of IgG to protein A binding sites is dependent mainly on the isoelectric point (PI). Below or above PI may cause elution of IgG from protein A (Wu et al 2011). Since a complete virus is antigenic and broken particles in the vaccine cannot potentially cause immunogenicity in the animals, a complete virion, such as the 146S fraction of FMDV, should be present in the vaccine.…”

Section: Introductionmentioning

confidence: 99%

Immuno-affinity chromatography for purification of IgG from hyper-immune sera raised against 146S fraction of Foot and Mouth Disease Virus for diagnostic purposes

Munir,

Anjum,

Altaf

et al. 2023

Polish Journal of Veterinary Sciences

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Immunoaffinity chromatography (IAC) is a fundamental isolation and purification tool which is incorporated in a substantial range of therapeutic and diagnostic applications. This study has reappraised the usefulness of immunoaffinity chromatography for the purification of polyclonal antibodies. Protein A based IAC is a convenient and reliable method for purification of IgG, from hyperimmunesera (HIS) raised in experimental animals such as rabbits, guinea pigs and mice to be utilized in pharmaceutics and diagnostics. The 146S fraction of Foot and Mouth Disease virus (FMDV) TCID50=10 5.6 was cultured on a baby hamster kidney cell line 21 (BHK-21), concentrated using salt precipitation method using PEG 6000, purified by size exclusion chromatography (SEC) using Sepharose-30 at 254nm absorbance. Purification of 146S FMDV was analyzed using 12% SDS-PAGE which provided two bands of light and heavy chains. The alum-based vaccine, consisting of ≥10µg of 146S FMDV, was applied in 10 male rabbits and 10 male guinea pigs and two animals of each group were taken as a negative control. The titer of serum was calculated using virus neutralization test. A Protein-A kit (Thermo scientific-44667, 0528.2) was used to purify HIS raised against 146S FMDV and validated using 12% SDS PAGE in reducing condition. The data demonstrate that protein-A affinity chromatography is an efficient tool for the purification of antibodies from hyper-immune sera raised against 146S FMDV and can be used for the production of diagnostic kits e.g. Enzyme linked immuno-sorbent assay (ELISA) and radioimmunoassay.

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Flexible Automation System for Parameter Screening and Small-Scale Protein Purification Applied to Colostrum Samples

Bach,

Palent,

Zastrow

et al. 2024

2024 IEEE 20th International Conference on Automation Science and Engineering (CASE)

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Isolation and Purification of Bioactive Proteins from Bovine Colostrum

Cited by 3 publications

References 40 publications

Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat

Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat

Immuno-affinity chromatography for purification of IgG from hyper-immune sera raised against 146S fraction of Foot and Mouth Disease Virus for diagnostic purposes

Flexible Automation System for Parameter Screening and Small-Scale Protein Purification Applied to Colostrum Samples

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