Engineering High Affinity Superantigens by Phage Display

Enever, Carolyn; Tomlinson, Ian M.; Lund, John; Levens, Michaela; Holliger, Philipp

doi:10.1016/j.jmb.2005.01.020

Cited by 18 publications

(11 citation statements)

References 35 publications

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“…Unlike Protein A, Protein G binds polyclonal rat IgG, human IgG3 and mouse IgG1 [99]. Protein A binds to some VH-3 subclass heavy chains [100]. Protein G exhibits CH1 interaction [101].…”

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

“…Proteins A, G, and L all bind to different target sites in conserved framework regions of antibodies, without compromising an antibody's ability to interact with its antigen [100]. Protein A-based affinity media are most frequently used to purify full-size antibodies because they recognize targets possessing the Fc domain, however antibody fragments typically lack the Fc region which is why Protein A resins are of limited use in Ab-fragment purification (see Table 1).…”

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

“…Genetic engineering could possibly allow Protein L to expand its versatility and specificity (see below). As the binding site for Protein L resides in framework region 1 of the variable domain of kappa light chain subtypes (kappa I, III, and IV from humans and kappa I and V from mice) [94,96,110], fragments derived from antibodies that have kappa light chain subtypes can be purified using Protein L [100]. Since Protein L interacts with the light chain, it has no immunoglobulin class restrictions and offers the potential of being a "broadly if not generally" useful affinity ligand [109].…”

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

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Antibody Fragments and Their Purification by Protein L Affinity Chromatography

Rodrigo¹,

Gruvegard²,

Alstine

2015

Antibodies

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Antibodies and related proteins comprise one of the largest and fastest-growing classes of protein pharmaceuticals. A majority of such molecules are monoclonal antibodies; however, many new entities are antibody fragments. Due to their structural, physiological, and pharmacological properties, antibody fragments offer new biopharmaceutical opportunities. In the case of recombinant full-length antibodies with suitable Fc regions, two or three column purification processes centered around Protein A affinity chromatography have proven to be fast, efficient, robust, cost-effective, and scalable. Most antibody fragments lack Fc and suitable affinity for Protein A. Adapting proven antibody purification processes to antibody fragments demands different affinity chromatography. Such technology must offer the unit operation advantages noted above, and be suitable for most of the many different types of antibody fragments. Protein L affinity chromatography appears to fulfill these criteria-suggesting its consideration as a key unit operation in antibody fragment processing.

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“…Unlike Protein A, Protein G binds polyclonal rat IgG, human IgG3 and mouse IgG1 [99]. Protein A binds to some VH-3 subclass heavy chains [100]. Protein G exhibits CH1 interaction [101].…”

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

Section: Protein L and Its Use In Antibody Fragment Bioprocessingmentioning

confidence: 99%

See 1 more Smart Citation

Antibody Fragments and Their Purification by Protein L Affinity Chromatography

Rodrigo¹,

Gruvegard²,

Alstine

2015

Antibodies

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“…The PCR products were cloned into pIT2 vectors (Enever et al, 2005) and 60-80 colonies were selected to confirm inserted genes by PCR screening using primer set for second PCR (Table 1). Approximately 20 samples of each group were analyzed by sequencing (MWG, Germany).…”

Section: Sequence Analysismentioning

confidence: 99%

Overexpression and unique rearrangement of VH2 transcripts in immunoglobulin variable heavy chain genes in ankylosing spondylitis patients

Kim

Lee

et al. 2010

Exp Mol Med

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“…Phage display provides the selection and identification of peptides binding to a variety of proteins by screening of extremely large and diverse libraries of DNA-encoded peptides (Sidhu et al, 2003). Although phage display techniques have been used for numerous biological studies such as designing high affinity super antigens (Enever et al, 2005), mapping functional binding sites (Sidhu et al, 2003), and identification of peptide binders to inorganic materials (Sarikaya et al, 2003), the greatest interest has been aroused for development of peptide ligands as recognition reagents for a specific protein target.…”

Section: Introductionmentioning

confidence: 99%

Thermodynamic and structural analysis of interactions between peptide ligands and SEB

Dudak¹,

Soykut²,

Oğuz³

et al. 2010

J of Molecular Recognition

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Staphylococcal enterotoxin B (SEB) is an exotoxin produced by Staphylococcus aureus and commonly associated with food poisoning. In this study, SEB-binding peptides were identified by screening a phage displayed peptide library. The binding of peptides to SEB was tested with isothermal titration calorimetry (ITC) and of the five selected peptides, three showed affinity to SEB, with one measured to have the highest affinity constant (10(5) M(-1)). ITC revealed that the interaction of peptide ligands with SEB was driven entropically and the binding was dominated by hydrophobic interactions. Circular dichroism (CD) measurements and molecular dynamics (MD) simulations, together, give a structural insight into the interaction of peptides with SEB. While SEB binding peptides showed random coil structure before binding, after complex formation they had more ordered structures. The peptide with highest affinity to SEB showed stable conformation during MD simulation. Taken together, our approach about thermodynamic and structural characterization of peptide ligands can be used to develop aptamers, with high affinity and selectivity, for biosensor applications.

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Engineering High Affinity Superantigens by Phage Display

Cited by 18 publications

References 35 publications

Antibody Fragments and Their Purification by Protein L Affinity Chromatography

Antibody Fragments and Their Purification by Protein L Affinity Chromatography

Overexpression and unique rearrangement of VH2 transcripts in immunoglobulin variable heavy chain genes in ankylosing spondylitis patients

Thermodynamic and structural analysis of interactions between peptide ligands and SEB

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