Epitope Mapping of Monoclonal Antibodies to Keratin 19 Using Keratin Fragments, Synthetic Peptides and Phage Peptide Libraries

Böttger, Volker; Stasiak, Pauline C.; Harrison, Dolores L.; Mellerick, Dervla M.; Lane, E. Birgitte

doi:10.1111/j.1432-1033.1995.tb20721.x

Cited by 32 publications

(19 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Monoclonal antibodies used in this study were: LL001 against K14 (Purkis et al, 1990); E3 against K17 (Guelstein et al, 1988), LP34 reacting with keratins K1, K5, K6, and K18 (Lane and Alexander, 1990), and LP2K against K19 (Bö ttger et al, 1995). A rabbit polyclonal antiserum, RbaK5 (BL18), was used against K5 (Purkis et al, 1990).…”

Section: Keratin Expressionmentioning

confidence: 99%

Keratin 14-Null Cells as a Model to Test the Efficacy of Gene Therapy Approaches in Epithelial Cells

D’Alessandro

Coats

Jonkmann

et al. 2011

Journal of Investigative Dermatology

View full text Add to dashboard Cite

Skin fragility disorders caused by keratin mutations are incurable, and a better understanding of their etiology is needed to find new ways to improve and treat these conditions. The best-studied skin fragility disorder is epidermolysis bullosa simplex (EBS), an autosomal dominant condition caused by mutations in keratin 5 (K5) or K14. To analyze disease mechanisms and develop gene therapy strategies, we have used keratinocyte cell lines derived from EBS patients as model systems. Here, we describe two cell lines established from EBS patients with K14-null mutations. We analyze the responses of these cells to stress assays previously shown to discriminate between wild-type and keratin-mutant keratinocytes, to directly evaluate the efficacy of rescuing K14-null cells by supplementation with wild-type K14 complementary DNA (cDNA). The K14-null cells show elevated levels of stress correlating with reduced normal keratin function. By transfecting wild-type K14 into these cells, we demonstrate "proof of principle" that an add-back approach can significantly rescue the normal keratinocyte behavior profile. These K14-null cell lines provide a disease model for studying the effects of keratin ablation in EBS patients and to test the efficacy of gene add-back and other therapy approaches in keratinocytes.

show abstract

Section: Keratin Expressionmentioning

confidence: 99%

Keratin 14-Null Cells as a Model to Test the Efficacy of Gene Therapy Approaches in Epithelial Cells

D’Alessandro

Coats

Jonkmann

et al. 2011

Journal of Investigative Dermatology

View full text Add to dashboard Cite

show abstract

“…As evident in literature, phage sequences are selected by MAbs based on their affinity to the given monoclonal. (53,54) Thus the six amino acids although essential for binding to MAb, it is possible that the intervening amino acids in the native protein could play a critical role in its immunogeneicity. It is also possible that amino acids distant in the primary sequence but adjacent in the tertiary structure of the protein may play a critical role in modulating the immune response.…”

Section: Discussionmentioning

confidence: 99%

Selection of an Immunogenic and Protective Epitope of the PsaA Protein ofStreptococcus pneumoniaeUsing a Phage Display Library

Srivastava¹,

Zeiler²,

Smithson³

et al. 2000

Hybridoma

View full text Add to dashboard Cite

Streptococcus pneumoniae is an important pathogen that causes disease in young and elderly individuals. The currently available polysaccharide vaccines have limited efficacy in those age groups most susceptible to pneumococcal infections. This study focuses on mapping the epitopes of a surface protein of S. pneumoniae by biopanning a 15 mer phage display library using 5 different monoclonal antibodies (MAbs) against the Pneumoccal surface adhesin A (PsaA). PsaA is a component of the bacterial cell wall that is highly species specific and is involved in bacterial adherence and virulence. Biopanning of the phage display library reveals three distinct epitopes on the PsaA protein. The sequence homology of these epitopes ranges from two to six amino acids when compared to the native PsaA protein type 2. Two of these epitopes have been evaluated for their immunogeneicity in mice. The peptide selected by the MAbs 8G12, 6F6, and 1B7 is referred to as the consensus peptide and is immunogenic in mice. Optimal anti-PsaA response is observed in mice immunized with 50microg of the consensus peptide complexed to proteosomes in 1:1 ratio. The anti-PsaA response is significantly lower than the response to the PsaA native protein. The peptide selected by monoclonal antibody 4E9 in its lipidated form is significantly protective in mice challenged with S. pneumoniae serotype 2 when compared to mice immunized with the native protein. These results show that the selected epitopes of PsaA protein are immunogenic and protective in mice. These epitopes need to be evaluated further as alternatives to currently available vaccines.

show abstract

“…In addition, these homologous amino acids did not necessarily form a continuous motif. Sequences chosen from a phage display library are selected by monoclonal antibodies on the basis of their affinity to the given monoclonal antibody [41,42]. Although these homologous amino acids are essential for binding to monoclonal antibody, it is possible that the intervening amino acids in the native protein could play a critical role in its immunogenicity.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Pneumococcal Carriage in Mice by Subcutaneous Immunization with Peptides from the Common Surface Protein Pneumococcal Surface Adhesin A

Johnson¹,

Dykes²,

Jue

et al. 2002

J INFECT DIS

View full text Add to dashboard Cite

Pneumococcal surface adhesin A (PsaA), a common protein expressed on all 90 pneumococcal serotypes, is a vaccine candidate. Three anti-PsaA monoclonal antibody phage display-expressed monopeptides (15 mers), in various formulations as lipidated or nonlipidated multiantigenic peptides or as bi- or tripeptide constructs, were studied in a mouse nasopharyngeal carriage model to determine the inhibitory effect of induced antibodies on carriage of pneumococcal serotypes 2, 4, and 6B. Antibodies to each of the various peptides tested reduced carriage of the 3 serotypes. Reduction in carriage by nonlipidated multiantigenic peptide antibodies was highly variable (39%-94%; mean, 59%; standard deviation [SD], 20.2%); however, more-consistent results were observed in mice immunized with lipidated (56%-98%; mean, 69%; SD, 13.6%) and combination or bipeptide (55%-91%; mean, 76%; SD, 13.1%) formulations. These peptides are immunogenic, and their induced antibodies reduce carriage in mice. PsaA peptides demonstrate potential for being important new vaccines against pneumococcal carriage, otitis media, and invasive pneumococcal disease.

show abstract

Epitope Mapping of Monoclonal Antibodies to Keratin 19 Using Keratin Fragments, Synthetic Peptides and Phage Peptide Libraries

Cited by 32 publications

References 33 publications

Keratin 14-Null Cells as a Model to Test the Efficacy of Gene Therapy Approaches in Epithelial Cells

Keratin 14-Null Cells as a Model to Test the Efficacy of Gene Therapy Approaches in Epithelial Cells

Selection of an Immunogenic and Protective Epitope of the PsaA Protein ofStreptococcus pneumoniaeUsing a Phage Display Library

Inhibition of Pneumococcal Carriage in Mice by Subcutaneous Immunization with Peptides from the Common Surface Protein Pneumococcal Surface Adhesin A

Contact Info

Product

Resources

About