The enzymes that are used in gene manipulation are described in Chapter 5. The discovery of restriction endonucleases, and the use of type II restriction enzymes in generating DNA fragments is outlined. The three types of fragment ends (blunt or flush-ended, 5’ protruding and 3’ protruding) enable DNA from different sources to be joined together, using the enzyme DNA ligase. DNA modifying enzymes include polymerases (DNA and RNA polymerases and reverse transcriptase), nucleases that act on the ends of molecules (exonucleases) or within a DNA strand (endonucleases), transferases, kinases and phosphatases. The enzymes make up the ‘toolkit’ that allows DNA to be manipulated in the test tube (in vitro) to generate recombinant molecules.

Bioinformatics is discussed in Chapter 11. The complex nature of the subject and its interaction with other disciplines are outlined, and the inter-dependence of bioinformatics, the development of computer hardware and the internet is stressed. The nature and range of biological databases are outlined, from the inception of nucleic acid databases in the 1970s to the present breadth of primary and secondary databases that are repositories for information on nucleic acid and protein sequences, interactions between cellular components, biochemical pathways, pharmacological targets and many other data sets derived from existing information. Genome sequence databases are used to illustrate the tools needed to assemble, collate, annotate and interrogate the data, and the impact of bioinformatics in enabling experiments and protocols to to be conducted in silico is discussed.

To introduce the subject, the history of genetics since Mendel’s work which was rediscovered in 1900 is outlined. The discovery of the structure of DNA in 1953 marked the start of the molecular genetics era. When restriction enzymes and DNA ligase were discovered, DNA fragments could be cut and joined, with the first recombinant DNA molecules generated in 1972. Rapid methods for sequencing DNA were developed in the late 1970s and eventually were improved to the level needed to enable the Human Genome Project to be undertaken. The completion of this in 2003 marked the start of the ‘post-genomic era’ that led to further development of the technology and a reduction in time and cost of genome sequencing. We are now firmly in the post-genomic era, where DNA technology is having a major impact in areas such as transgenic plants and animals, genome editing, diagnosis and treatment of disease, forensic analysis and personalised medicine.

In Chapter 13, what is needed to analyse cloned genes, and how this can be achieved are considered. The ultimate structural information is the sequence of the gene, and thus DNA sequencing has become a standard part of any cloning experiment. Although genome sequencing has led to a greater emphasis on bioinformatics-based analysis, methods such as restriction mapping, gel mobility shift assays, DNA footprinting and the various blotting techniques are still needed to confirm and link structure and function. The yeast hybrid systems have become important for analysing DNA∼protein, RNA∼protein and protein∼protein interactions, and DNA microarray technology and its extensions have changed the way in which gene expression is investigated. High-throughput analysis at genome and transcriptome levels is now routine and cost-effective. Genome projects have now generated vast amounts of sequence data, and the fields of comparative genomics and structural genomics are well established. Sequencing large numbers of genomes has now become possible and is leading to new discoveries and therapeutic interventions based on genome analysis.

Medical and forensic applications of recombinant DNA are described in Chapter 15. The range of genetically based diseases is outlined, and potential therapies discussed, covering diagnosis of infection, comparative genomics, development of vaccines, therapeutic antibodies and xenotransplantation. Treatment using gene therapy approaches is described, and the relatively limited success of gene therapy is considered in the context of its initial promise and the expectations that emerged from this. RNA-based therapies are covered by discussing RNA interference and antisense oligonucleotides, and the medical applications of genome editing are considered. The CCR5 controversy, known as the ‘CRISPR babies scandal’, is mentioned as an example of how the overall system can fail to prevent unethical practices when these are driven by determined scientists and clinicians. DNA profiling for analysis of DNA is described, and its use in forensic, legal and other applications is outlined.

Chapter 4 describes how living systems are organised at the molecular level, beginning with the chemistry of carbon-based systems and the concept of emergent properties. The genetic code and the flow of information are introduced as a key central theme, and the structure of DNA and RNA is presented. An outline of gene structure and organisation in prokaryotes and eukaryotes is followed by the description of transcription and translation as the mechanisms by which genes are expressed. A broader look at how genomes are organised leads to an outline of the transcriptome and proteome as two important concepts that are key to understanding how the genome functions in adaptive and developmental contexts.

New product development processes need to be compliant to regulatory requirements, and this chapter highlights the salient processes and quality systems to put into place to achieve success. Project management is made simple with specific tools provided here. Customer feedback is channeled into specific product characteristics, and the right tools are shown in this chapter. The biopharma industry has statistics showing less than 10% of starting compounds succeed in reaching market approval, and this chapter explains what causes these failures. The key issues that have repeatedly caused failure during device and diagnostic product development are also pointed out. Ethical decisions have to be made during product development as shown in this chapter. Outsourcing is a real option due to the availability of many contract research and manufacturing organizations, and judicious use of this option is discussed in this chapter. Key milestones that reduce risk and show transition from early stage to preclinical prototype stages are reviewed here. Does the popular concept of minimum viable product in software development apply in biomedicine prototyping? Other similar questions that help the reader understand pitfalls and best practices are answered here.

Conducting market research to find solutions, identifying opportunities and defining the value of new inventions are some of the key points covered in this chapter. A carefully defined indication can make the difference between success and failure in medical product development and this chapter explains how to get better at nailing the exact problem to be solved. Market segmentation examples and cases show how to prevent being misled on market size and market projections. A referral chain tool is presented for closely analyzing market positioning and value proposition of the new technology or product. Key market drivers and hurdles used to dynamically determine market size, adoption rates, and strategize on product development cycles are discussed and presented in this chapter.

How do you read a patent and what subject matter is patentable? What is the purpose of a patent? Who is an inventor on the patent if work is done by many people on the project? What is the process of obtaining a patent in my country and globally? Read this chapter to see how you could lose commercialization rights to your own invention. When exactly does an invention or idea become patentable? Once you own a patent, how can you make money from it? What is the process of licensing and the key terms that should be negotiated in such a license agreement? What is the use of a copyright or a trade secret in biotech? What exactly constitutes patent infringement ? These questions and many others are addressed in this chapter on intellectual property.

This chapter simplifies the complex multi-payer healthcare reimbursement market and explains how to position your product for successful reimbursement. The best time to bring reimbursement planning into the product development process is discussed here. The U.S. healthcare system is used as a baseline and the healthcare systems of other countries are reviewed briefly. Reimbursement for devices and administered drugs is based on many factors, and this chapter shows the steps a biomedical product company can take to maximize revenues in the US Healthcare system. The basics of reimbursement – coverage, coding, and payment – are explained in simple terms with diagrams. Case studies help show how individual companies have addressed the reimbursement process for novel breakthrough technologies.

Steps taken to start a new venture can make for rocky road ahead if consideration is not given to the points reviewed in this chapter. How to select and build a team and fairly distribute the founder’s equity, how to select an advisory board or a board of directors, and the importance of establishing a culture within the new company are all points discussed in detail and highlighted through personal stories and case examples. The main components of a business plan are covered in many texts and blogs, so this chapter focuses on the practical issues that few academic texts discuss, such as: how to perform due diligence on your investors and tips on creating slide decks , pitching and presenting business plans, and structuring financials and milestone to meet investors key concerns. The sources of financing and expectations of investors are reviewed with a view to guiding the entrepreneur or executive through the key elements for success, including successful closing on a term sheet or preparing for due diligence so that the process moves smoothly towards closure of the financing. The specific challenges facing an academic technopreneur moving into a decision-making executive (CSO or CEO) role are reviewed and guidance offered on utilizing the strength of the team around them.

The vast world of biotechnology applications to human health is reviewed and the terminology used in the rest of the book is defined here. An overview of the industry, the value chains, the specific types of human health products covered in this text are presented in this chapter. A time-tested way to analyze an industry’s attractiveness for new entrants is presented here using Porter’s five forces model. Technology trends such as mobile health, artificial intelligence, 3D printing, cell and gene therapy, and robotics are presented to the reader in the context of the mission of improving human health. The overall process of development of new products in these various segments of drugs, devices and diagnostics sectors is reviewed here. The reader will leave this chapter with a 30,000-foot view of the industry dynamics and understand the context within which product commercialization is to be done.