Alexion Pharmaceuticals, Inc.

Shana Frederickson

Katherine S Bowdish

Mark Renshaw

Toshiaki Maruyama

John R McWhirter

Ying-Chi Lin

Martha Wild

Maruyama Toshiaki

Antibodies or fragments thereof having CDR regions replaced or fused with biologically active peptides are described. Flanking sequences may optionally be attached at one or both the carboxy-terminal and amino-terminal ends of the peptide in covalent association with adjacent framework regions. Compositions containing such antibodies or fragments thereof are useful in therapeutic and diagnostic modalities.

Rationally designed antibodies

Antibodies or fragments thereof having at least two CDR regions replaced or fused with biologically active peptides are described. Compositions containing such antibodies or fragments thereof are useful in therapeutic and diagnostic modalities.

Phagemid vectors containing a sequence of features between a Col E1 origin and an f1 origin are useful for display of polypeptides or proteins, including antibody libraries.

Phagemid vectors

Templates that are engineered to contain a predetermined sequence and a hairpin structure are provided by a nested oligonucleotide extension reaction. The engineered template allows Single Primer Amplification (SPA) to amplify a target sequence within the engineered template. In particularly useful embodiments, the target sequences from the engineered templates are cloned into expression vehicles to provide a library of polypeptides or proteins, such as, for example, an antibody library.

Nested oligonucleotides containing a hairpin for nucleic acid amplification

Methods of amplifying nucleic acid have now been discovered which include the steps of: a) annealing a primer to a template nucleic acid sequence, the primer having a first portion which anneals to the template and a second portion of predetermined sequence; b) synthesizing a polynucleotide that anneals to and is complementary to the portion of the template between the location at which the first portion of the primer anneals to the template and the end of the template, the polynucleotide having a first end and a second end, wherein the first end incorporates the primer; c) separating the polynucleotide synthesized in step (b) from the template; d) annealing a nested oligonucleotide to the second end of the polynucleotide synthesized in step (b), the nested oligonucleotide having a first portion that anneals to the second end of the polynucleotide and a second portion having the same predetermined sequence as the second portion of the primer; e) extending the polynucleotide synthesized in step (b) to provide a terminal portion thereof that is complementary to the predetermined sequence; and f) amplifying the extended polynucleotide using a single primer having the predetermined sequence. In particularly useful embodiments, the methods are used to amplify a repertoire of IgA antibodies.

Engineered templates and their use in single primer amplification

Methods of amplifying nucleic acid have now been discovered which include the steps of: a) annealing a primer to a template nucleic acid sequence, the primer having a first portion which anneals to the template and a second portion of predetermined sequence; b) synthesizing a polynucleotide that anneals to and is complementary to the portion of the template between the location at which the first portion of the primer anneals to the template and the end of the template, the polynucleotide having a first end and a second end, wherein the first end incorporates the primer; c) separating the polynucleotide synthesized in step (b) from the template; d) annealing a nested oligonucleotide to the second end of the polynucleotide synthesized in step (b), the nested oligonucleotide having a first portion that anneals to the second end of the polynucleotide and a second portion having the same predetermined sequence as the second portion of the primer; e) extending the polynucleotide synthesized in step (b) to provide a terminal portion thereof that is complementary to the predetermined sequence; and f) amplifying the extended polynucleotide using a single primer having the predetermined sequence.

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Solana Beach, CA, United States of America

Shana Frederickson