Bubble extraction and affinity chromatography techniques for human trypsin inhibitors enrichment and purification

Elena Sbrana, Maria Spinetti, Ferdinando Secco, Giorgio Raspi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Bubble extraction combined with affinity chromatography can be considered as fully reliable techniques to concentrate and separate proteins and enzymes, and could be applied to the treatment of large samples to purify analytes from diluted solutions, on a preparative scale. In order to assess the possibility of employment of these techniques for the preconcentration of proteins in biological fluids, they have been applied to isolate and purify Kazal-type and Kunitz-type trypsin inhibitors which are usually present in human urine in very low concentration and of renowned biomedical interest, especially for their anti-metastatic activity. A three-step procedure, including direct RP-HPLC monitoring of the inhibitors at intermediate and final steps, is described here. Recoveries obtained are in the range 70-90%, and the highly purified inhibitors display a fully retained trypsin-inhibiting activity.

Original languageEnglish (US)
Pages (from-to)1475-1479
Number of pages5
JournalPolyhedron
Volume21
Issue number14-15
DOIs
StatePublished - Jun 15 2002
Externally publishedYes

Fingerprint

Affinity chromatography
trypsin
Trypsin Inhibitors
chromatography
Affinity Chromatography
purification
inhibitors
Purification
affinity
bubbles
Proteins
Trypsin
proteins
Enzymes
urine
High Pressure Liquid Chromatography
Urine
Recovery
Fluids
enzymes

Keywords

  • Affinity chromatography
  • Bubble extraction
  • RP-HPLC
  • Urinary trypsin inhibitors

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry
  • Physical and Theoretical Chemistry
  • Materials Chemistry

Cite this

Bubble extraction and affinity chromatography techniques for human trypsin inhibitors enrichment and purification. / Sbrana, Elena; Spinetti, Maria; Secco, Ferdinando; Raspi, Giorgio.

In: Polyhedron, Vol. 21, No. 14-15, 15.06.2002, p. 1475-1479.

Research output: Contribution to journalArticle

Sbrana, Elena ; Spinetti, Maria ; Secco, Ferdinando ; Raspi, Giorgio. / Bubble extraction and affinity chromatography techniques for human trypsin inhibitors enrichment and purification. In: Polyhedron. 2002 ; Vol. 21, No. 14-15. pp. 1475-1479.
@article{a276a31392a848a6a8bb394e08c37722,
title = "Bubble extraction and affinity chromatography techniques for human trypsin inhibitors enrichment and purification",
abstract = "Bubble extraction combined with affinity chromatography can be considered as fully reliable techniques to concentrate and separate proteins and enzymes, and could be applied to the treatment of large samples to purify analytes from diluted solutions, on a preparative scale. In order to assess the possibility of employment of these techniques for the preconcentration of proteins in biological fluids, they have been applied to isolate and purify Kazal-type and Kunitz-type trypsin inhibitors which are usually present in human urine in very low concentration and of renowned biomedical interest, especially for their anti-metastatic activity. A three-step procedure, including direct RP-HPLC monitoring of the inhibitors at intermediate and final steps, is described here. Recoveries obtained are in the range 70-90{\%}, and the highly purified inhibitors display a fully retained trypsin-inhibiting activity.",
keywords = "Affinity chromatography, Bubble extraction, RP-HPLC, Urinary trypsin inhibitors",
author = "Elena Sbrana and Maria Spinetti and Ferdinando Secco and Giorgio Raspi",
year = "2002",
month = "6",
day = "15",
doi = "10.1016/S0277-5387(02)00966-X",
language = "English (US)",
volume = "21",
pages = "1475--1479",
journal = "Polyhedron",
issn = "0277-5387",
publisher = "Elsevier Limited",
number = "14-15",

}

TY - JOUR

T1 - Bubble extraction and affinity chromatography techniques for human trypsin inhibitors enrichment and purification

AU - Sbrana, Elena

AU - Spinetti, Maria

AU - Secco, Ferdinando

AU - Raspi, Giorgio

PY - 2002/6/15

Y1 - 2002/6/15

N2 - Bubble extraction combined with affinity chromatography can be considered as fully reliable techniques to concentrate and separate proteins and enzymes, and could be applied to the treatment of large samples to purify analytes from diluted solutions, on a preparative scale. In order to assess the possibility of employment of these techniques for the preconcentration of proteins in biological fluids, they have been applied to isolate and purify Kazal-type and Kunitz-type trypsin inhibitors which are usually present in human urine in very low concentration and of renowned biomedical interest, especially for their anti-metastatic activity. A three-step procedure, including direct RP-HPLC monitoring of the inhibitors at intermediate and final steps, is described here. Recoveries obtained are in the range 70-90%, and the highly purified inhibitors display a fully retained trypsin-inhibiting activity.

AB - Bubble extraction combined with affinity chromatography can be considered as fully reliable techniques to concentrate and separate proteins and enzymes, and could be applied to the treatment of large samples to purify analytes from diluted solutions, on a preparative scale. In order to assess the possibility of employment of these techniques for the preconcentration of proteins in biological fluids, they have been applied to isolate and purify Kazal-type and Kunitz-type trypsin inhibitors which are usually present in human urine in very low concentration and of renowned biomedical interest, especially for their anti-metastatic activity. A three-step procedure, including direct RP-HPLC monitoring of the inhibitors at intermediate and final steps, is described here. Recoveries obtained are in the range 70-90%, and the highly purified inhibitors display a fully retained trypsin-inhibiting activity.

KW - Affinity chromatography

KW - Bubble extraction

KW - RP-HPLC

KW - Urinary trypsin inhibitors

UR - http://www.scopus.com/inward/record.url?scp=0037098220&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037098220&partnerID=8YFLogxK

U2 - 10.1016/S0277-5387(02)00966-X

DO - 10.1016/S0277-5387(02)00966-X

M3 - Article

AN - SCOPUS:0037098220

VL - 21

SP - 1475

EP - 1479

JO - Polyhedron

JF - Polyhedron

SN - 0277-5387

IS - 14-15

ER -