Selection and characterization of lipase abzyme from phage displayed antibody libraries. 2007

Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
Department of Chemistry, National Dong Hwa University, Hualien 974, Taiwan, ROC.

Antibodies with enzymatic activity were named abzymes or catalytic antibodies. In the present study, the lipolytic abzymes were selected from the phage displayed antibody libraries against a transition state analog (TSA) of lipases/esterases. After three rounds of selection, four monoclonal phage particles capable of binding significantly with the TSA were obtained. The soluble scFv antibody fragments were further expressed and obtained using Escherichia coli strain HB2151. The binding capabilities and the apparent enzymatic activities of the purified antibody proteins were measured. The 3D structures of the expressed antibodies were also predicted through homology modeling and binding-site prediction algorithm. The present method demonstrates that selection from phage displayed antibody libraries is an efficient and convenient means to find new abzymes.

UI MeSH Term Description Entries
D007135 Immunoglobulin Variable Region That region of the immunoglobulin molecule that varies in its amino acid sequence and composition, and comprises the binding site for a specific antigen. It is located at the N-terminus of the Fab fragment of the immunoglobulin. It includes hypervariable regions (COMPLEMENTARITY DETERMINING REGIONS) and framework regions. Variable Region, Ig,Variable Region, Immunoglobulin,Framework Region, Immunoglobulin,Fv Antibody Fragments,Fv Fragments,Ig Framework Region,Ig Variable Region,Immunoglobulin Framework Region,Immunoglobulin Fv Fragments,Immunoglobulin V,Antibody Fragment, Fv,Antibody Fragments, Fv,Fragment, Fv,Fragment, Fv Antibody,Fragment, Immunoglobulin Fv,Fragments, Fv,Fragments, Fv Antibody,Fragments, Immunoglobulin Fv,Framework Region, Ig,Framework Regions, Ig,Framework Regions, Immunoglobulin,Fv Antibody Fragment,Fv Fragment,Fv Fragment, Immunoglobulin,Fv Fragments, Immunoglobulin,Ig Framework Regions,Ig Variable Regions,Immunoglobulin Framework Regions,Immunoglobulin Fv Fragment,Immunoglobulin Variable Regions,Regions, Immunoglobulin Variable,Variable Regions, Ig,Variable Regions, Immunoglobulin
D008049 Lipase An enzyme of the hydrolase class that catalyzes the reaction of triacylglycerol and water to yield diacylglycerol and a fatty acid anion. It is produced by glands on the tongue and by the pancreas and initiates the digestion of dietary fats. (From Dorland, 27th ed) EC 3.1.1.3. Triacylglycerol Lipase,Tributyrinase,Triglyceride Lipase,Acid Lipase,Acid Lipase A,Acid Lipase B,Acid Lipase I,Acid Lipase II,Exolipase,Monoester Lipase,Triacylglycerol Hydrolase,Triglyceridase,Triolean Hydrolase,Hydrolase, Triacylglycerol,Hydrolase, Triolean,Lipase A, Acid,Lipase B, Acid,Lipase I, Acid,Lipase II, Acid,Lipase, Acid,Lipase, Monoester,Lipase, Triglyceride
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D000465 Algorithms A procedure consisting of a sequence of algebraic formulas and/or logical steps to calculate or determine a given task. Algorithm
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001665 Binding Sites The parts of a macromolecule that directly participate in its specific combination with another molecule. Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining
D016415 Sequence Alignment The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms. Sequence Homology Determination,Determination, Sequence Homology,Alignment, Sequence,Alignments, Sequence,Determinations, Sequence Homology,Sequence Alignments,Sequence Homology Determinations
D017151 Antibodies, Catalytic Antibodies that can catalyze a wide variety of chemical reactions. They are characterized by high substrate specificity and share many mechanistic features with enzymes. Catalytic Antibodies,Antibody, Catalytic,Catalytic Antibody

Related Publications

Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
January 2015, Journal of visualized experiments : JoVE,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
October 1996, Journal of immunological methods,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
August 2015, Biochemical and biophysical research communications,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
January 1998, Methods in molecular medicine,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
June 2001, Journal of molecular biology,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
April 1995, Gene,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
February 1999, Combinatorial chemistry & high throughput screening,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
August 1998, Current opinion in biotechnology,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
February 1998, Biochemical Society transactions,
Max K Leong, and Chinpiao Chen, and Ker-Chang Shar, and David Shiuan
January 2015, Methods in molecular biology (Clifton, N.J.),
Copied contents to your clipboard!