Pacific Biosciences
Pacific Biosciences of California, Inc. (aka PacBio) is an American biotechnology company founded in 2004 that develops and manufactures systems for gene sequencing and some novel real time biological observation. PacBio describes its platform as single-molecule real-time sequencing (SMRT), based on the properties of zero-mode waveguides. History. The company was founded based on research done at Cornell University that combined semiconductor processing and photonics with biotechnology research. Three graduate students in the lab of Professors Watt W. Webb — Jonas Korlach — and Harold Craighead — Steve Turner and Mathieu Foquet — became the first employees. It began under  the name Nanofluidics, Inc. The company raised nearly in six rounds of primarily venture capital financing, making it one of the most capitalized startups in 2010 leading up to their public offering in October of that year. Key investors included Mohr Davidow Ventures, Kleiner, Perkins, Caufield & Byers, Alloy Ventures, and Wellcome Trust. The company's first commercial product, the PacBio RS, was sold to a limited set of customers in 2010 and was commercially released in early 2011. A subsequent version of the sequencer called the PacBio RS II was released in April 2013. On September 25, 2013, a partnership  between Pacific Biosciences and Roche Diagnostics was announced for the development of in vitro diagnostic products using the technology, with Roche providing in the deal. In 2015, the company launched a new sequencing instrument called the Sequel System with approximately 7-fold greater capacity than the PacBio RS II. Then in April 2019 the company launched the Sequel II system with a further 8-fold increase in throughput. In October 2020 the company announced the Sequel IIe, which added onboard consensus computation to the sequencing instrument so DNA sequences produced are already corrected for accuracy. In February 2021, the company  announced a $900M investment in convertible senior notes by SoftBank Group. In July 2021 in a stock offering, the company raised $300M and announced the intent to acquire the short-read sequencing startup Omniome for $800M in cash and stock. Then in August, it acquired sample preparation company Circulomics. Leadership. In 2004, Kleiner Perkin's entrepreneur-in-residence Hugh Martin became CEO. On 6 January 2012 board member Michael Hunkapiller, PhD assumed the role of CEO. Hunkapiller retired in September 2020, and was replaced by chairman of the board Christian Henry. Henry was a  former executive VP and chief commercial officer of Illumina before joining the board at PacBio in July 2018. Going public. In an initial public offering on October 27, 2010, Pacific Biosciences sold 12,500,000 shares at a price of $16 per share and raised approximately $200 million. The shares trade on NASDAQ under the symbol PACB. Illumina. On 1 November 2018, Illumina, Inc. agreed to purchase PacBio for US$1.2 billion in cash. The deal was expected to close in the fourth quarter of 2019. In December 2019, the Federal Trade Commission sued to block the acquisition. The deal  was abandoned with an announcement on 2 January 2020. Illumina further agreed to pay Pacific Biosciences a $98 million US termination fee plus previously agreed upon deal extension payments of $22 million US in February and $6 million US in March 2020. Recognitions. The company received the Technology Pioneer Award from the World Economic Forum in 2009. In 2010, "The Scientist" named the company and their first product the top life science innovation of the year, and the company received the 2010 Advanced Sequencing Technology Award from the National Human Genome Research Institute. "Technology Review" magazine included  them in their list of the top 50 most innovative companies for both 2010 and 2011. Founder and Chief Technical Officer Dr. Stephen Turner was awarded the 2010 Ewing Marion Kauffman Foundation Outstanding Postdoctoral Entrepreneur award for his work at the company. Products. Sequencing instruments. The company's first scientific instrument, called the “PacBio RS”, was released to a limited set of eleven customers in late 2010. Sequencing provider GATC Biotech was selected by Pacific Biosciences as its first European service provider in late 2010. The product was then commercially released in early 2011. A new version of the sequencer  called the "PacBio RS II" was released in April 2013; it produced longer sequence reads and offered higher throughput than the original RS instrument. The RS instrument will officially be supported until the end of 2021. In September 2015, the company released a new sequencing instrument, the Sequel System. The sequencer has increased capacity with 1 million zero-mode waveguides compared to 150,000 in the PacBio RS II, and is approximately one-third the size and one-half the price of the PacBio RS II. In April 2019, the company released an upgraded Sequel II system with support for a  new SMRT Cell with eight million ZMW's, increasing the expected throughput per SMRT Cell by a factor of eight. In October 2020, the company announced the Sequel IIe system which adds additional compute capacity to the Sequel II system. With that compute all DNA reads produced by the system are automatically corrected for accuracy. Reagents and SMRT Cells. To use either instrument, customers must also purchase reagent packs for DNA preparation and sequencing and small consumables called “SMRT Cells”. Cells for the RS sequencer are slightly less than one-centimeter square and contains tens of thousands of zero-mode  waveguides. Cells for the Sequel sequencer are about 2.5 cm square and contains one million zero-mode waveguides, whereas cells for the Sequel II sequencer contain eight million zero-mode waveguides. The cells for the RS sequencer are sold in packs of eight. Cells for the Sequel or Sequel II sequencers are sold in packs of four. On 19 September 2018, PacBio released the Sequel 6.0 software and 3.0 chemistry. Performance differs between large-insert libraries and high molecular weight DNA versus shorter-insert libraries below ~15,000 bases in length. For larger templates, average read lengths are up to 30,000  bases. For shorter-insert libraries, the average read length is up to 100,000 bases while reading the same molecule in a circle. The latter shorter-insert libraries then yield up to 50 billion bases from a single SMRT® Cell. On 1 October 2019, PacBio released the 8.0 software and 2.0 chemistry for Sequel II. For larger templates read as "continuous long reads", an example human library yielded N50 read length of 52,456 and yield per cell is 182 GB. For libraries below ~20,000 bases, read in circular consensus sequencing, yield per cell is quoted at 450 GB or about  30 GB of HiFi corrected reads. Software and Applications. Their secondary analysis bioinformatics product for the RS, called “SMRT Analysis”, was open source. For the Sequel system the secondary analysis software was reorganized as the "SMRT Link" application. In 2013, the company released new bioinformatics tools for automated genome assembly (HGAP) and finishing to 99.999% accuracy (Quiver). Key publications. Before the first commercial release of their sequencer, scientists published in January 2009 the first sequence data generated from a single-molecule real-time sequencing in the journal Science. Then in April 2010, scientists published a paper in Nature showing that  they had used zero-mode waveguides to perform real-time observation of ribosomal translation. Demonstrating the value for bacterial sequencing, scientists from Pacific Biosciences and other institutions published in January 2011 a paper in the New England Journal of Medicine demonstrating the origin of the 2010 cholera outbreak in Haiti. In August 2011, Pacific Biosciences scientists and collaborators at other organizations published a paper in the New England Journal of Medicine describing the classification of the E. coli strain causing the virulent responsible for hundreds of cases of hemolytic–uremic syndrome. This paper showed that the strain of E. coli  responsible for the outbreak had acquired a Shiga-toxin–encoding phage through lateral gene transfer. In July 2012, several papers were published in peer-review journals demonstrating methods to automate genome finishing for bacteria using single-molecule real-time sequencing. In 2013, scientists estimated that the majority of bacterial and archaeal genomes could be fully sequenced and assembled to closure using PacBio long reads. Several papers published by researchers at Pacific Biosciences demonstrated that the sequencing instrument can be used to collect data on methylation, DNA damage, and other epigenetic information. The polymerase that performs the sequencing reaction in the  zero-mode waveguides produces kinetic data that can be used to distinguish base modifications. In October 2012, scientists used SMRT sequencing to generate the methylomes of six bacteria, reporting their results in a paper in Nucleic Acids Research. With increasing read length and throughput, mammalian studies increased using the product. In April 2012, scientists from Pacific Biosciences, the University of California, and other institutes used SMRT sequencing to prove the validity of activating internal tandem duplication mutations in FLT3 as a therapeutic target in acute myeloid leukemia. Their findings were published in the journal Nature. In August 2012, scientists at  the Broad Institute published a paper reporting the findings of their evaluation of the Pacific Biosciences sequencer for SNP calling and discovery. Scientists reported in "Genome Research" in October 2012 the use of the PacBio platform to sequence the full repeat expansion in the FMR1 gene responsible for Fragile X Syndrome. A paper published in December 2012 offered the first demonstration of how to generate sequence data with the PacBio sequencer with no library preparation. In 2013, scientists published papers demonstrating the use of PacBio sequencing to analyze transcriptomes, showing that long reads were able to fully capture complete isoforms