Affinity binding of inclusion bodies on supermacroporous monolithic cryogels using labeling with specific antibodies

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Affinity binding of inclusion bodies on supermacroporous monolithic cryogels using labeling with specific antibodies

Ahlqvist, Josefin (author): Lund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Lund University, Lund, Sweden

Kumar, Ashok (author): Lund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Lund University, Lund, Sweden

Sundstrom, H. (author): Royal Institute of Technology, Stockholm, Sweden

Ledung, E. (author): Mälardalens högskola,Institutionen för biologi och kemiteknik

Hornsten, E. G. (author): SIK, Swedish Institute for Food and Biotechnology, Sweden

Enfors, Sven-Olof (author): Mälardalens högskola,KTH,Bioprocessteknik,Institutionen för biologi och kemiteknik

Mattiasson, B. (author): Lund University, Lund, Sweden

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Elsevier BV, 2006
2006
English.
In: Journal of Biotechnology. - : Elsevier BV. - 0168-1656 .- 1873-4863. ; 122:2, s. 216-225

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Journal article (peer-reviewed)

Abstract Subject headings

A new chromatographic method based on affinity supermacroporous monolithic cryogels is developed for binding and analyzing inclusion bodies during fermentation. The work demonstrated that it is possible to bind specific IgG and IgY antibodies to the 15 and 17 amino acids at the terminus ends of a 33 kDa target protein aggregated as inclusion bodies. The antibody treated inclusion bodies from lysed fermentation broth can be specifically retained in protein A and pseudo-biospecific ligand sulfamethazine modified supermacroporous cryogels. The degree of binding of IgG and IgY treated inclusion bodies to the Protein A and sulfamethazine gels are investigated, as well as the influence of pH on the sulfamethazine ligand. Optimum binding of 78 and 72% was observed on both protein A and sulfamethazine modified cryogel columns, respectively, using IgG labeling of the inclusion bodies. The antibody treated inclusion bodies pass through unretained in the sulfamethazine supermacroporous gel at pH that does not favour the binding between the ligand on the gel and the antibodies on the surface of inclusion bodies. Also the unlabeled inclusion bodies went through the gel unretained, showing no non-specific binding or trapping within the gel. These findings may very well be the foundation for the building of a powerful analytical tool during fermentation of inclusion bodies as well as a convenient way to purify them from fermentation broth. These results also support our earlier findings [Kumar, A., Plieva, F.M., Galaev, I.Yu., Mattiasson, B.. 2003. Affinity fractionation of lymphocytes using a monolithic cyogel. J. Immunol. Methods 283, 185-194] with mammalian cells that were surface labeled with specific antibodies and recognized on protein A supermacroporous gels. A general binding and separation system can be established on antibody binding cryogel affinity matrices.

Subject headings

TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
LANTBRUKSVETENSKAPER -- Lantbruksvetenskap, skogsbruk och fiske -- Livsmedelsvetenskap (hsv//swe)
AGRICULTURAL SCIENCES -- Agriculture, Forestry and Fisheries -- Food Science (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology (hsv//eng)

Keyword

antibodies
IgG
IgY
protein A
sulfamethazine
cryogels
inclusion bodies
escherichia-coli
protein
chromatography
expression
cells
bodies
inclusion
cryogels
sulfamethazine
protein A
IgY
antibodies
IgG

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ref (subject category)
art (subject category)

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ENGINEERING AND TECHNOLOGY: ENGINEERING AND ...; and Industrial Biote ...

AGRICULTURAL SCIENCES: AGRICULTURAL SCI ...; and Agriculture Fore ...; and Food Science

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Medical Biotechn ...

Articles in the publication: Journal of Biote ...

By the university: Royal Institute of Technology; Lund University; RISE; Mälardalen University

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