Co- and Post-Translational Modifications of Therapeutic Antibodies and Proteins

T. Shantha Raju

• Publisher: Wiley
• Year: 2019
• Print ISBN:
  9781119053316
• eBook ISBN:
  9781119053361
• Edition: 1
• Topic: Sciences biologiques
• Subtopic: Biochimie

A Comprehensive Guide to Crucial Attributes of Therapeutic Proteins in Biological Pharmaceuticals

With this book, Dr. Raju offers a valuable resource for professionals involved in research and development of biopharmaceutical and biosimilar drugs. This is a highly relevant work, as medical practitioners have increasingly turned to biopharmaceutical medicines in their search for safe and reliable treatments for complex diseases, while pharmaceutical researchers seek to expand the availability of biopharmaceuticals and create more affordable biosimilar alternatives. 

Readers receive a thorough overview of the major co-translational modifications (CTMs) and post-translational modifications (PTMs) of therapeutic proteins relevant to the development of biotherapeutics. The majority of chapters detail individual CTMs and PTMs that may affect the physicochemical, biochemical, biological, pharmacokinetic, immunological, toxicological etc. properties of proteins. In addition, readers are guided on the methodology necessary to analyze and characterize these modifications. Thus, readers gain not only an understanding of CTMs/PTMs, but also the ability to design and assess their own structure-function studies for experimental molecules. Specific features and topics include: 

• Discussion of the research behind and expansion of biopharmaceuticals
• Twenty chapters detailing relevant CTMs and PTMs of proteins, such as glycosylation, oxidation, phosphorylation, methylation, proteolysis, etc.
• Each chapter offers an introduction and guide to the mechanisms and biological significance of an individual CTM or PTM, including practical guidance for experiment design and analysis
• An appendix of biologic pharmaceuticals currently on the market, along with an assessment of their PTMs and overall safety and efficacy

This volume will prove a key reference on the shelves of industry and academic researchers involved in the study and development of biochemistry, molecular biology, biopharmaceuticals and proteins in medicine, particularly as biopharmaceuticals and biosimilars become ever more prominent tools in the field of healthcare.

1
Introduction to Co‐ and Post‐translational Modifications of Proteins

Proteins are mostly biologically derived macromolecules containing amino acid residues as their constituent monomers (Marrack and Hoch 1949; Zamecnik 1950). Amino acids are small organic molecules containing amine and carboxylic acid as their functional groups (Sakami and Harrington 1963). The amine group of one amino acid can interact with the carboxylic acid group of another amino acid or another molecule of the same amino acid to form a stable covalent bond (Hertweck 2011). Such covalent bonds between amine groups and carboxylic acid groups of amino acids are often referred to as amide bonds. These amide bonds are also called as peptide bonds. Peptides are mostly linear chain molecules containing two or more amino acid residues that are linked by peptide bonds (Work 1961). The peptide bonds can form extended polypeptide chains resulting in very high molecular weight peptides. Such very high molecular weight peptides are also referred to as proteins. Proteins, unlike most peptides, mostly contain highly ordered structures and are most commonly synthesized by biological processes within a given biological system or organism. However, recently cell‐free in vitro synthesis systems are also established using enzymes and/or chemicals to produce proteins (Perez et al. 2016). Such systems are usually referred to as cell‐free protein synthesis (CFPS) systems (Semenchenko et al. 2016; Jia et al. 2017).

Figure 1.1 Chemical structure of 20 different amino acid residues commonly found in human proteins.

Biosynthesis of proteins involves the ribosomes in which mRNA translate the nascent polypeptide chains. During the biosynthesis of polypeptide chains amino acids, available within a given cell and/or organism, are used to construct amide/peptide bonds. There are atleast 20 different amino acid residues that exist in nature and several more of their derivatives are also present in living organisms. The chemical structure of 20 different amino acids are shown in Figure 1.1. Physicochemical properties along with the three and one letter abbreviations of these amino acids and some of their derivatives are shown in Table 1.1. In living organisms, about 20 different amino acids are directly translated by RNA that are represented by the triple letter codons in the genetic code during the biosynthesis of the nascent polypeptide chains (Wu et al. 2013). Upon biosynthesis, the polypeptide chain undergoes different processes to form the active protein, which ultimately might become structural proteins, functional proteins, enzymes, receptors, transporters, translocators, proteases, binding proteins, antibodies, cytokines, etc.

Table 1.1 Amino acid residues and their functionality.

Names (in alphabetic order)	Three letter abbreviations	One letter abbreviations	Nature of functionality	Functional group	Comments
Alanine	Ala	A	Neutral	Aliphatic	Nonessential
Arginine	Arg	R	Basic	Amine	Semiessential
Asparagine	Asn	N	Polar	Amide	Nonessential
Aspartic acid	Asp	D	Acidic	Carboxylic acid	Nonessential
Aspartate or asparagine	Asx	B	Acidic or polar	Carboxylic acid or amide	Nonessential
Cysteine	Cys	C	Nucleophilic	Thiol	Nonessential
Glutamate	Glu	E	Polar	Amide	Nonessential
Glutamine	Gln	Q	Polar	Amide	Nonessential
Glutamine or glutamate	Glx	Z	Polar	Amide	Nonessential
Glycine	Gly	G	Neutral	Proteogenic	Nonessential
Histidine	His	H	Basic	Reactive heterocyclic ring	Nonessential
Isoleucine	Ile	I	Hydrophobic	Aliphatic	Essential
Leucine	Leu	L	Hydrophobic	Aliphatic	Essential
Lysine	Lys	K	Basic	Amine	Essential
Methionine	Met	M	Hydrophobic	Aliphatic	Essential
Phenylalanine	Phe	F	Hydrophobic	Aromatic	Essential
Proline	Pro	P	Hydrophobic	Ring structure	Nonessential
Pyrrolysine	Pyl	O	Basic	Pyrroline	Nonessential
Selenocysteine	Sec	U	Neutral	Selenol	Nonessential
Serine	Ser	S	Polar	Hydroxyl	Nonessential
Threonine	Thr	T	Polar	Hydroxyl	Essential
Tryptophan	Trp	W	Hydrophobic	Aromatic	Essential
Tyrosine	Tyr	Y	Hydrophobic	Aromatic	Nonessential
Valine	Val	V	Hydrophobic	Aliphatic	Essential

The monomer...

Table of contents

• Cover
• Table of Contents
• Preface
• About the Author
• Abbreviations
• 1 Introduction to Co‐ and Post‐translational Modifications of Proteins
• 2 Acetylation of Proteins
• 3 C‐terminal Lys or Arg Clipping in Proteins
• 4 Cysteinylation of Proteins
• 5 Deamidation of Proteins
• 6 Glycation of Proteins
• 7 Glycosylation of Proteins
• 8 N‐glycosylation of Proteins
• 9 O‐glycosylation of Proteins
• 10 Hydroxylation of Proteins
• 11 Methylation of Proteins
• 12 Oxidation of Proteins
• 13 Phosphorylation of Proteins
• 14 Prenylation of Proteins
• 15 Proteolysis of Proteins
• 16 Selenylation
• 17 Signal Peptides
• 18 Sulfation of Proteins and Glycoproteins
• 19 SUMOylation
• 20 Ubiquitination
• 21 Other CTMs and PTMs of Proteins
• Appendix A
• Index
• End User License Agreement