Advances in Bio-instrumentation

Booklet published from work at VG Biotech
There's no doubt that since, say 1980, the development and application of scientific instrumentation in the life sciences has been rapid, enabling and has surpassed the expectations of even those in the field. I list some of these below:

  • Mass spectrometers: Electrospray and Matrix-assisted laser desorption time-of-flight
  • Protein sequencing: Gas phase
  • Gene sequencing: Automated Sanger and Oxford nanopore sequencers
  • Microscopy: cryoelectron microscopy, confocal microscopy, atomic force microscopy and others

I participated in the early development of electrospray mass spectrometry (ESMS) as applied to proteins and peptides, working with FISONS Instruments (VG Biotech) at their Altrincham labs near Manchester. Acting as a consultant to my former employer (Delta Biotechnology) I was also able to use ESMS to look at the quality of recombinant proteins like HSA.

I produced a pair of illustrative booklets, the most significant of which explored the application of ESMS to determine protein post-translational processing. Prior to this, candidate proteins had to be digested to provide lower molecular weight peptides which then needed to be separated by chromatography in many instances.

ESMS is especially valuable with intact glycoproteins for detecting and identifying "glycoforms". When combined with MS/MS methods and sometimes enzymatic digest, complete molecular descriptions of glycoproteins can be realised. We are approaching whole cell glycoproteomics, if we're not already there as I write this page.

Protein identification Mass spectrometry now is an important technology in proteomics. Peptide mass fingerprinting (of protein mixtures in the case of proteomics analysis) enables searching mass spectral databases of predicted protein fragment masses. This can be refined further by performing MS/MS analysis to obtain sequence information on selected peptides.

I was very fortunate that my host institution agreed to purchase a "gas phase" protein sequencer for my lab on the understanding that I would assist other institute groups with sequence analysis (my equipment request then was rather like asking for an MRI scanner now!). We had the second instrument installed in the UK. It enabled me to compare sequences from annexins (as mentioned elsewhere).

Gas phase sequencing technology was developed in the US at Cal Tech in LeRoy Hood's lab with Mike Hunkapiller. The spin-out business was known as Applied Biosystems. The sequencer versions gradually improved until I was using a fully automated instrument at Delta Biotechnology Ltd in Nottingham to sequence recombinant proteins under development as therapeutics.

As expected the capability of protein sequencing technology has continued to develop.As with the fluorescent dye-labelling methods applied to sequence nucleic acids, analogous labelling techniques are now being developed to sequence proteins. The eventually may lead to "super-proteomics" approaches that can determine protein sequences in isolation or in mixtures.

Gas phase protein sequencer supplied by ABI biosystems

Recombinant therapeutic protein stability in formulations I undertook a contract for Thermometric Ltd, A UK business that manufactures extremely sensitive microcalorimeters.

Heat flow examined in recombinant HSA samples held at 57 deg C in a Thermometric microcalorimeter

I gained this contact at one of the trade fairs I organised alongside my POPE conference. I am grateful to Nottingham University for allowing me to run my experiments in their labs. Protein (or small molecule) therapeutics can change chemically or degrade in other ways during storage in the delivery ampoules. A highly topical example would be the mRNA COVID-19 vaccines formulated in lipid micelles or liposomes.

The thermometric equipment can pick this up as enthalpy changes (heat absorption or emission) with incredible sensitivity. For the company I used their instrument to look at recombinant HSA in solution. The figure shows the heat changes of excipient (buffer) and recombinant HSA with and without added stabilizers at 57oC over 20 hours. I was highly impressed with this equipment which picked up changes even in therapeutics in solid tablet form.