Phage display as a tool to study human autoantibodies and autoantigens in systemic autoimmune disease. Selection of recombinant (auto)-antibodies specific for human autoantigens in rheumatic disease (RA, SLE, SSc) from human autoimmune-patient and immunized chicken derived phage display libraries

Background: Phage display is a biotechnological technique that presents peptides with coated proteins on the surface of phage. In the last two decades, growing applications of phage display in various fields of biotechnology have been investigated. Phage display libraries allow to present billions of peptides on phage surface for selection of a specific peptide with the desired affinity. Objective: In this regard, high-affinity phage antibodies against tumor antigens are produced and applied for diagnosis and treatment of cancer. Method: Moreover, phage display libraries are employed to select the high affinity T Cell Receptors (TCRs) for the peptide-MHC complex which is an attractive approach in cancer immunotherapy. Due to immunogenic properties of phage particles, phage-based vaccines do not require adjuvant, in addition the phage particles can effectively take up by Antigen Presenting Cells (APCs). Results: Taken together, phage-based cancer vaccines are ideal candidates that provide a key for eradication of tumor cells. Conclusion: In this review, we focus on various applications of a phage display platform in different types of cancer immunotherapy approaches.

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Phage display and kinetic selection of antibodies that specifically inhibit amyloid self-replication

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1700407114 ◽

2017 ◽

Vol 114 (25) ◽

pp. 6444-6449 ◽

Cited By ~ 32

Author(s):

Anna Munke ◽

Jonas Persson ◽

Tanja Weiffert ◽

Erwin De Genst ◽

Georg Meisl ◽

...

Keyword(s):

Phage Display ◽

Amyloid Fibrils ◽

Amyloid Β ◽

Amyloid Β Peptide ◽

Nucleation Process ◽

Single Chain ◽

Secondary Nucleation ◽

Single Chain Antibody ◽

Phage Display Libraries ◽

Selection Of

The aggregation of the amyloid β peptide (Aβ) into amyloid fibrils is a defining characteristic of Alzheimer’s disease. Because of the complexity of this aggregation process, effective therapeutic inhibitors will need to target the specific microscopic steps that lead to the production of neurotoxic species. We introduce a strategy for generating fibril-specific antibodies that selectively suppress fibril-dependent secondary nucleation of the 42-residue form of Aβ (Aβ42). We target this step because it has been shown to produce the majority of neurotoxic species during aggregation of Aβ42. Starting from large phage display libraries of single-chain antibody fragments (scFvs), the three-stage approach that we describe includes (i) selection of scFvs with high affinity for Aβ42 fibrils after removal of scFvs that bind Aβ42 in its monomeric form; (ii) ranking, by surface plasmon resonance affinity measurements, of the resulting candidate scFvs that bind to the Aβ42 fibrils; and (iii) kinetic screening and analysis to find the scFvs that inhibit selectively the fibril-catalyzed secondary nucleation process in Aβ42 aggregation. By applying this approach, we have identified four scFvs that inhibit specifically the fibril-dependent secondary nucleation process. Our method also makes it possible to discard antibodies that inhibit elongation, an important factor because the suppression of elongation does not target directly the production of toxic oligomers and may even lead to its increase. On the basis of our results, we suggest that the method described here could form the basis for rationally designed immunotherapy strategies to combat Alzheimer’s and related neurodegenerative diseases.

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Selection of high-affinity phage antibodies from phage display libraries

Nature Biotechnology ◽

10.1038/7959 ◽

1999 ◽

Vol 17 (4) ◽

pp. 397-399 ◽

Cited By ~ 67

Author(s):

Robert de Bruin ◽

Kees Spelt ◽

Joseph Mol ◽

Ronald Koes ◽

Francesca Quattrocchio

Keyword(s):

Phage Display ◽

High Affinity ◽

Phage Antibodies ◽

Phage Display Libraries ◽

Selection Of

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Strong selection of a few dominant CD8 clones in a TLR7-dependent autoimmune mouse model

10.1101/393819 ◽

2018 ◽

Cited By ~ 1

Author(s):

Peter A. Morawski ◽

Silvia Bolland

Keyword(s):

Autoimmune Disease ◽

Clonal Selection ◽

Specific Antigen ◽

Systemic Autoimmune Disease ◽

Cell Repertoire ◽

Cd8 Cells ◽

Nuclear Antigens ◽

Autoimmune Mouse ◽

Viral Responses ◽

Selection Of

Systemic lupus disease is characterized by the expansion of a self-reactive repertoire of B cells that, with the help of CD4 cells, generate IgG antibodies against common nuclear antigens. Meanwhile, the functional state and posible clonal selection of CD8 cells in lupus remain poorly defined. We previously described the activated but non-pathogenic phenotype of CD8+ T cells, some of which accumulate in the brain, in a model of systemic autoimmune disease triggered by increased copy number of the tlr7 gene (TLR7tg mice). Here we report, through the analysis of TCRβ sequences, that CD8+ cells from TLR7tg are strongly selected for a small number of clones, some of them reaching 30% of the repertoire, compared to less than 0.4% for a single top clone in wild type cells. High frequency clones are variable in sequence among individual TLR7tg mice and are distinct from top clones in WT, while cells from spleen and brain-resident cells from the same animals have perfect concordance. These results suggest that top CD8 clones are selected in stochastic fashion in each animal but limit further diversification, and that brain infiltrating CD8 cells in TLR7tg are not selected by a tissue specific antigen. This kind of extreme clonal dominance and narrowing of the CD8+ T cell repertoire coud potentially impair anti-viral responses and should be considered as an additional detrimental feature of chronic autoimmune disease.

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