Optimization of human serum albumin monoliths for chiral separations and high-performance affinity chromatography

Erika L. Pfaunmiller, Mahli Hartmann, Courtney M. Dupper, Sony Soman, David S. Hage

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Various organic-based monoliths were prepared and optimized for immobilization of the protein human serum albumin (HSA) as a binding agent for chiral separations and high-performance affinity chromatography. These monoliths contained co-polymers based on glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA) or GMA and trimethylolpropane trimethacrylate (TRIM). A mixture of cyclohexanol and 1-dodecanol was used as the porogen, with the ratio of these solvents being varied along with the polymerization temperature to generate a library of monoliths. These monoliths were used with both the Schiff base and epoxy immobilization methods and measured for their final content of HSA. Monoliths showing the highest protein content were further evaluated in chromatographic studies using R/S-warfarin and d/. l-tryptophan as model chiral solutes. A 2.6-2.7-fold increase in HSA content was obtained in the final monoliths when compared to similar HSA monoliths prepared according to the literature. The increased protein content made it possible for the new monoliths to provide higher retention and/or two-fold faster separations for the tested solutes when using 4.6 mm i.d. × 50 mm columns. These monoliths were also used to create 4.6 mm i.d. × 10 mm HSA microcolumns that could separate the same chiral solutes in only 1.5-6.0 min. The approaches used in this study could be extended to the separation of other chiral solutes and to the optimization of organic monoliths for use with additional proteins as binding agents.

Original languageEnglish (US)
Pages (from-to)198-207
Number of pages10
JournalJournal of Chromatography A
Volume1269
DOIs
StatePublished - Dec 21 2012

Fingerprint

Affinity chromatography
Affinity Chromatography
Serum Albumin
Immobilization
Proteins
Dodecanol
Cyclohexanols
Schiff Bases
Warfarin
Protein Binding
Tryptophan
Polymerization
Libraries
Polymers
Temperature

Keywords

  • Affinity monolith chromatography
  • Chiral separations
  • High-performance affinity chromatography
  • Human serum albumin
  • Monolith columns

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Optimization of human serum albumin monoliths for chiral separations and high-performance affinity chromatography. / Pfaunmiller, Erika L.; Hartmann, Mahli; Dupper, Courtney M.; Soman, Sony; Hage, David S.

In: Journal of Chromatography A, Vol. 1269, 21.12.2012, p. 198-207.

Research output: Contribution to journalArticle

Pfaunmiller, Erika L. ; Hartmann, Mahli ; Dupper, Courtney M. ; Soman, Sony ; Hage, David S. / Optimization of human serum albumin monoliths for chiral separations and high-performance affinity chromatography. In: Journal of Chromatography A. 2012 ; Vol. 1269. pp. 198-207.
@article{4da329dde2d843b79d503061be2710c9,
title = "Optimization of human serum albumin monoliths for chiral separations and high-performance affinity chromatography",
abstract = "Various organic-based monoliths were prepared and optimized for immobilization of the protein human serum albumin (HSA) as a binding agent for chiral separations and high-performance affinity chromatography. These monoliths contained co-polymers based on glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA) or GMA and trimethylolpropane trimethacrylate (TRIM). A mixture of cyclohexanol and 1-dodecanol was used as the porogen, with the ratio of these solvents being varied along with the polymerization temperature to generate a library of monoliths. These monoliths were used with both the Schiff base and epoxy immobilization methods and measured for their final content of HSA. Monoliths showing the highest protein content were further evaluated in chromatographic studies using R/S-warfarin and d/. l-tryptophan as model chiral solutes. A 2.6-2.7-fold increase in HSA content was obtained in the final monoliths when compared to similar HSA monoliths prepared according to the literature. The increased protein content made it possible for the new monoliths to provide higher retention and/or two-fold faster separations for the tested solutes when using 4.6 mm i.d. × 50 mm columns. These monoliths were also used to create 4.6 mm i.d. × 10 mm HSA microcolumns that could separate the same chiral solutes in only 1.5-6.0 min. The approaches used in this study could be extended to the separation of other chiral solutes and to the optimization of organic monoliths for use with additional proteins as binding agents.",
keywords = "Affinity monolith chromatography, Chiral separations, High-performance affinity chromatography, Human serum albumin, Monolith columns",
author = "Pfaunmiller, {Erika L.} and Mahli Hartmann and Dupper, {Courtney M.} and Sony Soman and Hage, {David S.}",
year = "2012",
month = "12",
day = "21",
doi = "10.1016/j.chroma.2012.09.009",
language = "English (US)",
volume = "1269",
pages = "198--207",
journal = "Journal of Chromatography A",
issn = "0021-9673",

}

TY - JOUR

T1 - Optimization of human serum albumin monoliths for chiral separations and high-performance affinity chromatography

AU - Pfaunmiller, Erika L.

AU - Hartmann, Mahli

AU - Dupper, Courtney M.

AU - Soman, Sony

AU - Hage, David S.

PY - 2012/12/21

Y1 - 2012/12/21

N2 - Various organic-based monoliths were prepared and optimized for immobilization of the protein human serum albumin (HSA) as a binding agent for chiral separations and high-performance affinity chromatography. These monoliths contained co-polymers based on glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA) or GMA and trimethylolpropane trimethacrylate (TRIM). A mixture of cyclohexanol and 1-dodecanol was used as the porogen, with the ratio of these solvents being varied along with the polymerization temperature to generate a library of monoliths. These monoliths were used with both the Schiff base and epoxy immobilization methods and measured for their final content of HSA. Monoliths showing the highest protein content were further evaluated in chromatographic studies using R/S-warfarin and d/. l-tryptophan as model chiral solutes. A 2.6-2.7-fold increase in HSA content was obtained in the final monoliths when compared to similar HSA monoliths prepared according to the literature. The increased protein content made it possible for the new monoliths to provide higher retention and/or two-fold faster separations for the tested solutes when using 4.6 mm i.d. × 50 mm columns. These monoliths were also used to create 4.6 mm i.d. × 10 mm HSA microcolumns that could separate the same chiral solutes in only 1.5-6.0 min. The approaches used in this study could be extended to the separation of other chiral solutes and to the optimization of organic monoliths for use with additional proteins as binding agents.

AB - Various organic-based monoliths were prepared and optimized for immobilization of the protein human serum albumin (HSA) as a binding agent for chiral separations and high-performance affinity chromatography. These monoliths contained co-polymers based on glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA) or GMA and trimethylolpropane trimethacrylate (TRIM). A mixture of cyclohexanol and 1-dodecanol was used as the porogen, with the ratio of these solvents being varied along with the polymerization temperature to generate a library of monoliths. These monoliths were used with both the Schiff base and epoxy immobilization methods and measured for their final content of HSA. Monoliths showing the highest protein content were further evaluated in chromatographic studies using R/S-warfarin and d/. l-tryptophan as model chiral solutes. A 2.6-2.7-fold increase in HSA content was obtained in the final monoliths when compared to similar HSA monoliths prepared according to the literature. The increased protein content made it possible for the new monoliths to provide higher retention and/or two-fold faster separations for the tested solutes when using 4.6 mm i.d. × 50 mm columns. These monoliths were also used to create 4.6 mm i.d. × 10 mm HSA microcolumns that could separate the same chiral solutes in only 1.5-6.0 min. The approaches used in this study could be extended to the separation of other chiral solutes and to the optimization of organic monoliths for use with additional proteins as binding agents.

KW - Affinity monolith chromatography

KW - Chiral separations

KW - High-performance affinity chromatography

KW - Human serum albumin

KW - Monolith columns

UR - http://www.scopus.com/inward/record.url?scp=84870352291&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84870352291&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2012.09.009

DO - 10.1016/j.chroma.2012.09.009

M3 - Article

C2 - 23010249

AN - SCOPUS:84870352291

VL - 1269

SP - 198

EP - 207

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

ER -