Development of cross-linked antibody Fab fragment crystals for enantioselective separation of a drug enantiomer

Antti Vuolanto (Corresponding Author), Kristiina Kiviharju, Tarja Nevanen, Matti Leisola, Jouni Jokela

Research output: Contribution to journalArticleScientificpeer-review

12 Citations (Scopus)

Abstract

A novel immunoaffinity separation material, cross-linked antibody crystals (CLAC), was developed by crystallization and cross-linking of a recombinant antibody Fab fragment ENA5His capable of enantiospecific separation of a chiral drug, finrozole. Crystallization conditions of the antibody fragment were screened by hanging drop vapor diffusion experiments. The small-scale vapor diffusion crystallization was easily scaled up to 10-mL batch crystallization with a 70% yield. Also, the low affinity mutant of ENA5His having one single amino acid mutation crystallized at the same conditions. Simple glutaraldehyde cross-linking of the crystals yielded CLAC, which were insoluble in water, 5−100% methanol, and 2% DMSO in PBS buffer. Batch experiments showed that the CLAC material was functional as it specifically bound the desired enantiomer from the drug racemate.
Original languageEnglish
Pages (from-to)777-782
Number of pages6
JournalCrystal Growth and Design
Volume3
Issue number5
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Immunoglobulin Fab Fragments
Enantiomers
enantiomers
Crystallization
antibodies
Antibodies
drugs
fragments
Crystals
crystallization
Pharmaceutical Preparations
crystals
Vapors
Immunoglobulin Fragments
Functional materials
vapors
Glutaral
Dimethyl Sulfoxide
Methanol
mutations

Keywords

  • antibodies
  • antibody Fab'-fragment
  • antibody fragments
  • immobilization
  • crystallization

Cite this

Vuolanto, Antti ; Kiviharju, Kristiina ; Nevanen, Tarja ; Leisola, Matti ; Jokela, Jouni. / Development of cross-linked antibody Fab fragment crystals for enantioselective separation of a drug enantiomer. In: Crystal Growth and Design. 2003 ; Vol. 3, No. 5. pp. 777-782.
@article{ef36787eebde4b9fa3d59b77c06aa019,
title = "Development of cross-linked antibody Fab fragment crystals for enantioselective separation of a drug enantiomer",
abstract = "A novel immunoaffinity separation material, cross-linked antibody crystals (CLAC), was developed by crystallization and cross-linking of a recombinant antibody Fab fragment ENA5His capable of enantiospecific separation of a chiral drug, finrozole. Crystallization conditions of the antibody fragment were screened by hanging drop vapor diffusion experiments. The small-scale vapor diffusion crystallization was easily scaled up to 10-mL batch crystallization with a 70{\%} yield. Also, the low affinity mutant of ENA5His having one single amino acid mutation crystallized at the same conditions. Simple glutaraldehyde cross-linking of the crystals yielded CLAC, which were insoluble in water, 5−100{\%} methanol, and 2{\%} DMSO in PBS buffer. Batch experiments showed that the CLAC material was functional as it specifically bound the desired enantiomer from the drug racemate.",
keywords = "antibodies, antibody Fab'-fragment, antibody fragments, immobilization, crystallization",
author = "Antti Vuolanto and Kristiina Kiviharju and Tarja Nevanen and Matti Leisola and Jouni Jokela",
year = "2003",
doi = "10.1021/cg034021r",
language = "English",
volume = "3",
pages = "777--782",
journal = "Crystal Growth and Design",
issn = "1528-7483",
publisher = "American Chemical Society",
number = "5",

}

Development of cross-linked antibody Fab fragment crystals for enantioselective separation of a drug enantiomer. / Vuolanto, Antti (Corresponding Author); Kiviharju, Kristiina; Nevanen, Tarja; Leisola, Matti; Jokela, Jouni.

In: Crystal Growth and Design, Vol. 3, No. 5, 2003, p. 777-782.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Development of cross-linked antibody Fab fragment crystals for enantioselective separation of a drug enantiomer

AU - Vuolanto, Antti

AU - Kiviharju, Kristiina

AU - Nevanen, Tarja

AU - Leisola, Matti

AU - Jokela, Jouni

PY - 2003

Y1 - 2003

N2 - A novel immunoaffinity separation material, cross-linked antibody crystals (CLAC), was developed by crystallization and cross-linking of a recombinant antibody Fab fragment ENA5His capable of enantiospecific separation of a chiral drug, finrozole. Crystallization conditions of the antibody fragment were screened by hanging drop vapor diffusion experiments. The small-scale vapor diffusion crystallization was easily scaled up to 10-mL batch crystallization with a 70% yield. Also, the low affinity mutant of ENA5His having one single amino acid mutation crystallized at the same conditions. Simple glutaraldehyde cross-linking of the crystals yielded CLAC, which were insoluble in water, 5−100% methanol, and 2% DMSO in PBS buffer. Batch experiments showed that the CLAC material was functional as it specifically bound the desired enantiomer from the drug racemate.

AB - A novel immunoaffinity separation material, cross-linked antibody crystals (CLAC), was developed by crystallization and cross-linking of a recombinant antibody Fab fragment ENA5His capable of enantiospecific separation of a chiral drug, finrozole. Crystallization conditions of the antibody fragment were screened by hanging drop vapor diffusion experiments. The small-scale vapor diffusion crystallization was easily scaled up to 10-mL batch crystallization with a 70% yield. Also, the low affinity mutant of ENA5His having one single amino acid mutation crystallized at the same conditions. Simple glutaraldehyde cross-linking of the crystals yielded CLAC, which were insoluble in water, 5−100% methanol, and 2% DMSO in PBS buffer. Batch experiments showed that the CLAC material was functional as it specifically bound the desired enantiomer from the drug racemate.

KW - antibodies

KW - antibody Fab'-fragment

KW - antibody fragments

KW - immobilization

KW - crystallization

U2 - 10.1021/cg034021r

DO - 10.1021/cg034021r

M3 - Article

VL - 3

SP - 777

EP - 782

JO - Crystal Growth and Design

JF - Crystal Growth and Design

SN - 1528-7483

IS - 5

ER -