Phage Display of Cellulose Binding Domains for Biotechnological Application

2000 ◽  
pp. 168-189 ◽  
Author(s):  
Itai Benhar ◽  
Aviva Tamarkin ◽  
Lea Marash ◽  
Yevgeny Berdichevsky ◽  
Sima Yaron ◽  
...  
Keyword(s):  
Phage Display ◽  
Biotechnological Application ◽  
Binding Domains ◽  
Cellulose Binding

Combination of ribosome and phage display for fast selection of high affinity VHH antibody fragments

2019 ◽  
pp. 27-33
Author(s):  
YuE Kravchenko ◽  
SV Ivanov ◽  
DS Kravchenko ◽  
EI Frolova ◽  
SP Chumakov
Keyword(s):  
Phage Display ◽  
Selection Procedure ◽  
Free System ◽  
Phage Displays ◽  
High Affinity ◽  
Binding Domains ◽  
A Cell ◽  
Vhh Antibodies ◽  
Efficiency Of Selection ◽  
Selection Of

Selection of antibodies using phage display involves the preliminary cloning of the repertoire of sequences encoding antigen-binding domains into phagemid, which is considered the bottleneck of the method, limiting the resulting diversity of libraries and leading to the loss of poorly represented variants before the start of the selection procedure. Selection in cell-free conditions using a ribosomal display is devoid from this drawback, however is highly sensitive to PCR artifacts and the RNase contamination. The aim of the study was to test the efficiency of a combination of both methods, including pre-selection in a cell-free system to enrich the source library, followed by cloning and final selection using phage display. This approach may eliminate the shortcomings of each method and increase the efficiency of selection. For selection, alpaca VHH antibody sequences suitable for building an immune library were used due to the lack of VL domains. Analysis of immune libraries from the genes of the VH3, VHH3 and VH4 families showed that the VHH antibodies share in the VH3 and VH4 gene groups is insignificant, and selection from the combined library is less effective than from the VHH3 family of sequences. We found that the combination of ribosomal and phage displays leads to a higher enrichment of high-affinity fragments and avoids the loss of the original diversity during cloning. The combined method allowed us to obtain a greater number of different high-affinity sequences, and all the tested VHH fragments were able to specifically recognize the target, including the total protein extracts of cell cultures.

scholarly journals The type II and X cellulose-binding domains of Pseudomonas xylanase A potentiate catalytic activity against complex substrates by a common mechanism

1999 ◽  
Vol 342 (2) ◽  
pp. 473-480 ◽  
Author(s):  
Jaitinder GILL ◽  
Jane E. RIXON ◽  
David N. BOLAM ◽  
Simon MCQUEEN-MASON ◽  
Peter J. SIMPSON ◽  
...  
Keyword(s):  
Microcrystalline Cellulose ◽  
Plant Cell Wall ◽  
Catalytic Domain ◽  
Wall Material ◽  
Common Mechanism ◽  
Type Ii ◽  
Binding Domains ◽  
Covalently Linked ◽  
Cellulose Binding ◽  
Internal Domain

Xylanase A (Pf Xyn10A), in common with several other Pseudomonas fluorescens subsp. cellulosa polysaccharidases, consists of a Type II cellulose-binding domain (CBD), a catalytic domain (Pf Xyn10ACD) and an internal domain that exhibits homology to Type X CBDs. The Type X CBD of Pf Xyn10A, expressed as a discrete entity (CBDX) or fused to the catalytic domain (Pf Xyn10A′), bound to amorphous and bacterial microcrystalline cellulose with a Ka of 2.5×105 M-1. CBDX exhibited no affinity for soluble forms of cellulose or cello-oligosaccharides, suggesting that the domain interacts with multiple cellulose chains in the insoluble forms of the polysaccharide. Pf Xyn10A′ was 2-3 times more active against cellulose-hemicellulose complexes than Pf Xyn10ACD; however, Pf Xyn10A′ and Pf Xyn10ACD exhibited the same activity against soluble substrates. CBDX did not disrupt the structure of plant-cell-wall material or bacterial microcrystalline cellulose, and did not potentiate Pf Xyn10ACD when not covalently linked to the enzyme. There was no substantial difference in the affinity of full-length Pf Xyn10A and the enzyme's Type II CBD for cellulose. The activity of Pf Xyn10A against cellulose-hemicellulose complexes was similar to that of Pf Xyn10A′, and a derivative of Pf Xyn10A in which the Type II CBD is linked to the Pf Xyn10ACD via a serine-rich linker sequence [Bolam, Cireula, McQueen-Mason, Simpson, Williamson, Rixon, Boraston, Hazlewood and Gilbert (1998) Biochem J. 331, 775-781]. These data indicate that CBDX is functional in Pf Xyn10A and that no synergy, either in ligand binding or in the potentiation of catalysis, is evident between the Type II and X CBDs of the xylanase.

scholarly journals Generation of Metal-Binding Staphylococci through Surface Display of Combinatorially Engineered Cellulose-Binding Domains

2001 ◽  
Vol 67 (10) ◽  
pp. 4678-4684 ◽  
Author(s):  
Henrik Wernérus ◽  
Janne Lehtiö ◽  
Tuula Teeri ◽  
Per-Åke Nygren ◽  
Stefan Ståhl
Keyword(s):  
Metal Binding ◽  
Binding Capacity ◽  
Surface Display ◽  
Surface Proteins ◽  
Chimeric Proteins ◽  
Phage Display Technology ◽  
Binding Domains ◽  
Combinatorial Protein Engineering ◽  
Display Technology ◽  
Cellulose Binding

ABSTRACT Ni2+-binding staphylococci were generated through surface display of combinatorially engineered variants of a fungal cellulose-binding domain (CBD) from Trichoderma reeseicellulase Cel7A. Novel CBD variants were generated by combinatorial protein engineering through the randomization of 11 amino acid positions, and eight potentially Ni2+-binding CBDs were selected by phage display technology. These new variants were subsequently genetically introduced into chimeric surface proteins for surface display on Staphylococcus carnosus cells. The expressed chimeric proteins were shown to be properly targeted to the cell wall of S. carnosus cells, since full-length proteins could be extracted and affinity purified. Surface accessibility for the chimeric proteins was demonstrated, and furthermore, the engineered CBDs, now devoid of cellulose-binding capacity, were shown to be functional with regard to metal binding, since the recombinant staphylococci had gained Ni2+-binding capacity. Potential environmental applications for such tailor-made metal-binding bacteria as bioadsorbents in biofilters or biosensors are discussed.

scholarly journals Cellulose-Binding Domains as a Tool for Paper Recycling

2003 ◽  
pp. 105-115 ◽  
Author(s):  
H. Pala ◽  
R. Pinto ◽  
M. Mota ◽  
A. P. Duarte ◽  
F. M. Gama
Keyword(s):  
Paper Recycling ◽  
Binding Domains ◽  
Cellulose Binding

Biosynthesis of cellulose binding domains (CBDs)

2009 ◽  
Author(s):  
Jing Guo ◽  
Ming Tien ◽  
Jeffrey M Catchmark
Keyword(s):  
Binding Domains ◽  
Cellulose Binding

Identification of tandemly repeated type VI cellulose-binding domains in an endoglucanase from the aerobic soil bacterium Cellvibrio mixtus

1998 ◽  
Vol 49 (5) ◽  
pp. 552-559 ◽  
Author(s):  
C. M. G. A. Fontes ◽  
J. H. Clarke ◽  
G. P. Hazlewood ◽  
T. H. Fernandes ◽  
H. J. Gilbert ◽  
...  
Keyword(s):  
Soil Bacterium ◽  
Binding Domains ◽  
Cellulose Binding ◽  
Aerobic Soil
Sign in / Sign up

Export Citation Format

Share Document