Purification of Peg-Protein Conjugates by Centrifugal Precipitation Chromatography

Sookkumnerd, Terasut; Hsu, Jong-Ping; Ito, Yoichiro

doi:10.1081/jlc-100100466

Cited by 5 publications

(3 citation statements)

References 16 publications

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“…HPLC or SEC are highly useful for purification assuming that appropriate solvent systems and columns are available (59). Thermal cycling or enzyme recovery is commonly used when the polymer has a critical solution temperature.…”

Section: Design Of Protein-polymer Conjugatesmentioning

confidence: 99%

Protein‐Polymer Conjugates

Briand¹,

Kumar²,

Kasi³

2011

Encyclopedia of Polymer Science and Technology

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This chapter focuses on the design and applications of protein‐polymer conjugates. Protein‐polymer conjugates are synthesized by conjugating a polymer chain or many polymer chains onto a protein. The site of conjugation, protein, polymer, and stoichiometry are important criteria when designing a protein‐polymer conjugate. The important structural aspects and functions of proteins, such as hierarchical structure and activity, are discussed. The various methods of site‐specific conjugation, “grafting to”, “grafting from” and cofactor reconstitution, are included along the pros and cons of each method. Conjugates synthesized by random conjugation are also discussed. The specific reactions used for protein attachment are not included, as numerous synthetic reviews exist; however, a broad overview of the field is presented. Novel conjugate structures can be synthesized by attaching synthetic peptide sequences onto polymers. In addition to design considerations, the range of applications from drug delivery systems to biosensors are discussed. This chapter aims to aid in furthering the knowledge and growth of the synergistic combination of proteins and polymers.

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Section: Design Of Protein-polymer Conjugatesmentioning

confidence: 99%

Protein‐Polymer Conjugates

Briand¹,

Kumar²,

Kasi³

2011

Encyclopedia of Polymer Science and Technology

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“…The PEG-protein conjugates, however, are difficult to separate by the conventional chromatographic methods according to the number of PEG molecules per protein molecule. However, It was found that the present method was able to resolve each species as demonstrated by the separation of PEG-lysozyme conjugates using the standard separation method [7]. Fig.…”

Section: Applications Of Centrifugal Precipitation Chromatographymentioning

confidence: 99%

“…The method has been successfully applied to various kinds of biological samples including serum proteins [1-6], IgM [2,3] and IgG [3], recombinant ketosteroid isomerase [2-5], protein-PEG (polyethylene glycol) conjugate [3, 7] polysaccharide [3, 8, 9], polymerized pigments [10] plasmid DNA RNA and protein [11, 12], etc. In this review paper, this new method is introduced by describing the principle, instrumentation, a series of basic studies and three different types of applications including the standard separation, ligand-affinity separation, and immunoaffinity separation of proteins.…”

Section: Introductionmentioning

confidence: 99%

Centrifugal precipitation chromatography

Ito

Qiu

2010

Journal of Chromatography B

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Centrifugal precipitation chromatography separates analytes according their solubility in ammonium sulfate (AS) solution and other precipitants. The separation column is made from a pair of long spiral channels partitioned with a semipermeable membrane. In a typical separation, concentrated ammonium sulfate is eluted through one channel while water is eluted through the other channel in the opposite direction. The countercurrent process forms an exponential AS concentration gradient through the water channel. Consequently, protein samples injected into the water channel is subjected to a steadily increasing AS concentration and at the critical AS concentration they are precipitated and deposited in the channel bed by the centrifugal force. Then the chromatographic separation is started by gradually reducing the AS concentration in the AS channel which lowers the AS gradient concentration in the water channel. This results in dissolution of deposited proteins which are again precipitated at an advanced critical point as they move through the channel. Consequently, proteins repeat precipitation and dissolution through a long channel and finally eluted out from the column in the order of their solubility in the AS solution. The present method has been successfully applied to a number of analytes including human serum proteins, recombinant ketosteroid isomerase, carotenoid cleavage enzymes, plasmid DNA, polysaccharide, polymerized pigments, PEG-protein conjugates, etc. The method is capable to single out the target species of proteins by affinity ligand or immunoaffinity separation.

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Immunoaffinity centrifugal precipitation chromatography

Qiu

Ito

2007

Journal of Chromatography A

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Purification of Peg-Protein Conjugates by Centrifugal Precipitation Chromatography

Cited by 5 publications

References 16 publications

Protein‐Polymer Conjugates

Protein‐Polymer Conjugates

Centrifugal precipitation chromatography

Immunoaffinity centrifugal precipitation chromatography

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