Generation and Refinement of Peptide Mimetic Ligands for Paratope-Specific Purification of Monoclonal Antibodies

Murray, Andrea; Smith, Richard G.; Brady, Kevin; Williams, S. C.; Badley, R.A.; Price, Michael R.

doi:10.1006/abio.2001.5235

Cited by 10 publications

(8 citation statements)

References 32 publications

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“…These fragments are ideally suited for some therapeutic (Kontermann et al, 1997) and diagnostic Power and Hudson, 2000) applications where better tissue penetration and rapid clearance from the body are required, and can be used per se or reconstituted into a fully human antibody molecule. Affinity ligands binding specifically to the Fab fragments of immunoglobulins have been produced and comprise anti-antibodies raised against the immunoglobulin of interest (Huse et al, 2002), molecules interacting with CDR regions (antigen or epitope/mimotope peptides; Murray et al, 2001) or with constant framework moieties (PpL; Vola et al, 1995). However, biological affinity ligands are limited by high-cost of production and purification, limited life cycles and low scale-up potential (Lowe et al, 2001;Gulich et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L fromPeptostreptococcus magnus

Roque

Taipa²,

Lowe

2005

J. Mol. Recognit.

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Rational design and combinatorial chemistry were utilized to search for lead protein L (PpL) mimetics for application as affinity ligands for the purification of antibodies and small fragments, such as Fab and scFv, and as potential diagnostic or therapeutic agents. Inspection of the key structural features of the complex between PpL and human Fab prompted the de novo design and combinatorial synthesis of a 169-membered solid-phase ligand library, which was assessed for binding to human IgG and subsequent selectivity for the Fab fragment. Eight ligands were selected, chemically characterized and compared with a commercial PpL-adsorbent for binding pure immunoglobulin fractions. The most promising lead, ligand 8/7, when immobilized on an agarose support, behaved in a similar fashion to PpL in isolating Fab fragments from papain digests of human IgG to a final purity of 97%.

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Section: Discussionmentioning

confidence: 99%

Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L fromPeptostreptococcus magnus

Roque

Taipa²,

Lowe

2005

J. Mol. Recognit.

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“…It was found that these Mab's react with a peptide (PDTRPAP), which after coupling to Sepharose proved to be an efficient adsorbent for Mabrecovery, from either hybridoma tissue cultures (154,155) or ascites fluids (156). A peptide mimotope (mimicking a non-proteinaceous antigen) has been further generated by combined biological (phage display) and chemical combinatorial techniques for paratopespecific purification of Mab's (157).…”

Section: Peptidic Ligandsmentioning

confidence: 98%

Antibodies and Genetically Engineered Related Molecules: Production and Purification

Roque

Lowe

Taipa³

2004

Biotechnol. Prog.

328

208

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Antibodies and antibody derivatives constitute 20 % of biopharmaceutical products currently in development, and despite early failures of murine products, chimeric and humanized monoclonal antibodies are now viable therapeutics. A number of genetically engineered antibody constructions have emerged, including molecular hybrids or chimeras that can deliver a powerful toxin to a target such as a tumor cell. However, the general use in clinical practice of antibody therapeutics is dependent not only on the availability of products with required efficacy but also on the costs of therapy. As a rule, a significant percentage (50-80%) of the total manufacturing cost of a therapeutic antibody is incurred during downstream processing. The critical challenges posed by the production of novel antibody therapeutics include improving process economics and efficiency, to reduce costs, and fulfilling increasingly demanding quality criteria for Food and Drug Administration (FDA) approval. It is anticipated that novel affinity-based separations will emerge from the development of synthetic ligands tailored to specific biotechnological needs. These synthetic affinity ligands include peptides obtained by synthesis and screening of peptide combinatorial libraries and artificial non-peptidic ligands generated by a de novo process design and synthesis. The exceptional stability, improved selectivity, and low cost of these ligands can lead to more efficient, less expensive, and safer procedures for antibody purification at manufacturing scales. This review aims to highlight the current trends in the design and construction of genetically engineered antibodies and related molecules, the recombinant systems used for their production, and the development of novel affinity-based strategies for antibody recovery and purification.

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“…Although immunoassays are still the preferred choice [18], in cases where original antigens of the antibodies are not readily accessible, mimotopes can be used as surrogates. Mimotopes have been used for the development of new diagnostics [5, 19, 20], therapeutics [21–24], vaccines [25–28], and as ligands for affinity chromatography applications [29, 30]. In this context the MimoDB database, established to collect information and sequences derived from biopanning results of random libraries, lists more than 15000 target specific peptide sequences binding to more than 1300 target structures (http://immunet.cn/) [31].…”

Section: Discussionmentioning

confidence: 99%

Peptide microarrays enable rapid mimotope optimization for pharmacokinetic analysis of the novel therapeutic antibody IMAB362

Schnatbaum

Schmoldt

Daneschdar

et al. 2014

Biotechnology Journal

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As membrane proteins play an important role in a variety of life-threatening diseases, the development of therapeutic monoclonal antibodies against membrane proteins is of significant interest. Among many other requirements, the process of antibody drug development requires a set of tailor-made assays for the characterization of the antibodies and for monitoring their activity. Designing assays to characterize antibodies directed to membrane proteins is challenging, because the natural targets are often not available in a format that is compatible with a biochemical assay setup. Thus, alternatives that mimic the targeted membrane proteins are needed. In this study, we developed optimal peptidic mimotopes for the ELISA-based detection of the novel therapeutic antibody IMAB362 in biological samples. Initial hits were identified using phage display and these hits were optimized with the help of structure-activity relationship analysis on peptide microarrays. The optimized peptides showed binding constants in the low nanomolar to picomolar range, an improvement by a factor of up to 30 compared to the initial hits. The best mimotope (apparent KD = 0.15 nM) was successfully used for the ELISA-based quantification of IMAB362 in samples from a mouse pharmacokinetic study. The process described allows the rapid discovery of mimotopes for target proteins that are difficult to produce or handle, which can then be used in pre-clinical and clinical assays or for the purification of biological products.

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Generation and Refinement of Peptide Mimetic Ligands for Paratope-Specific Purification of Monoclonal Antibodies

Cited by 10 publications

References 32 publications

Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L fromPeptostreptococcus magnus

Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L fromPeptostreptococcus magnus

Antibodies and Genetically Engineered Related Molecules: Production and Purification

Peptide microarrays enable rapid mimotope optimization for pharmacokinetic analysis of the novel therapeutic antibody IMAB362

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