Peptide and Protein Engineering
From Concepts to Biotechnological Applications
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Protocol
Hybrid Silylated Peptides for the Design of Bio-functionalized Materials
Hybrid silylated peptides are useful starting compounds for the design of functionalized materials. Indeed, silylated peptides may react in soft conditions with other silylated species through Si–O–Si bonds, o...
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Phage Display Methodologies
In vitro selection of bacteriophages displaying specific protein binders from large combinatorial libraries is a well-established and very powerful technology. Therapeutic antibodies that have been evolved by ...
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Building Scarless Gene Libraries in the Chromosome of Bacteria
Genetic libraries of gene variants are frequently used in protein engineering or molecular evolution experiments. Expression libraries cloned in plasmids are relatively easy to prepare but have some limitation...
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Integrated Bacterial Production and Functional Screening of Expanded Cyclic Peptide Libraries for Identifying Chemical Rescuers of Pathogenic Protein Misfolding and Aggregation
Protein misfolding and aggregation are defining features of a wide range of human conditions, collectively termed protein misfolding diseases. These include disorders with diverse pathologies and symptoms, suc...
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SEA-Mediated Ligation Is Accelerated at Mildly Acidic pH: Application to the Formation of Difficult Peptide Junctions
The bis(2-sulfanylethyl)amido (SEA)-mediated ligation has been introduced in 2010 as a novel chemoselective peptide bond-forming reaction. SEA-mediated ligation is a useful reaction for protein total synthesis th...
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Using the Interactive Tool of the Protein Chemical Synthesis Database
Over the last 25 years, chemoselective amide bond-forming reactions have established themselves as an essential tool for the total chemical synthesis of peptides and proteins. This spectacular development is e...
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Fluoroglycoproteins by Copper-Free Strain-Promoted Azide–Alkyne Cycloaddition
This chapter describes a reliable two-step, metal-free protocol for the preparation of well-defined fluoroglycoproteins. It starts with a first alkylation step to chemoselectively install strained alkyne handl...
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Synthesis of Peptide-Oligoethylene Glycol (OEG) Conjugates for Multivalent Modification of Nanomaterials
Multivalent nanomaterials are designed to take advantage of the distinctive features that arise from the cooperativity of multiple “ligand-target” interactions, resulting in improved target affinities or enhan...
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CIS Display: DNA-Based Technology as a Platform for Discovery of Therapeutic Biologics
CIS display is a cell-free in vitro display technology, based on linear dsDNA templates. This platform allowed for the evolution of peptides, antibody fragments, and protein scaffolds, mainly for therapeutic a...
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Whole-Bacterium Ribosome Display Selection for Isolation of Antibacterial Affitins
Classically, in order to perform ribosome display selections, a randomized gene library encoding potential affinity proteins is incubated with a pure target of interest that is either immobilized on a solid su...
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Streptavidin (Sav)-Based Artificial Metalloenzymes: Cofactor Design Considerations and Large-Scale Expression of Host Protein Variants
The first part of the introduction presents artificial cofactors that are employed for the formation of streptavidin-based artificial metalloenzymes. Their interaction with the protein scaffold is illustrated ...
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Site-Specific Incorporation of Non-canonical Amino Acids by Amber Stop Codon Suppression in Escherichia coli
The site-specific incorporation of non-canonical amino acids with reactive side chains is a powerful tool for the directed chemical modification of proteins. Here we provide a protocol for the site-specific in...
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The Problem of Aspartimide Formation During Protein Chemical Synthesis Using SEA-Mediated Ligation
Aspartimide formation often complicates the solid-phase synthesis of peptides. Much less discussed is the potential occurrence of this side reaction during the coupling of peptide segments using chemoselective...
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Only a “Click” Away: Development of Arginine-Rich Peptide-Based Materials Using Click Chemistry
The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) coupling is one of the most interesting chemoselective reactions that match the “click chemistry” concept. Both azide and alkyne moieties, easily inco...
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An Orthogonal Fusion Tag for Efficient Protein Purification
In this chapter, we present an efficient method for stringent protein purification facilitated by a dual affinity tag referred to as ABDz1, which is based on a 5 kDa albumin-binding domain from Streptococcal Prot...
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Exploring Satellite DNAs: Specificities of Bivalve Mollusks Genomes
Noncoding DNA sequences repeated in tandem or satellite DNAs make an integral part of every eukaryotic genome. Development and application of new methodological approaches through time enabled gradual improvem...
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Bioactive Secondary Metabolites of Medicinal and Aromatic Plants and Their Disease-Fighting Properties
Medicinal and aromatic plants (MAPs) play a vital role in traditional as well as modern medicine. According to World Health Organization (WHO), approximately 80% of total world’s population still relies on anc...
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Synthesis and Evaluation of Dye-Ligand Affinity Adsorbents for Protein Purification
Dye-ligand affinity chromatography is a widely used technique in protein purification. The utility of the reactive dyes as affinity ligands results from their unique chemistry, which confers wide specificity t...
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The Genomics of Plant Satellite DNA
The twenty-first century began with a certain indifference to the research of satellite DNA (satDNA). Neither genome sequencing projects were able to accurately encompass the study of satDNA nor classic method...
