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Gary Keane

Antibody Production Manager

Institute of Science & the Environment

Contact Details

email: g.keane@worc.ac.uk

tel: 01905 855204

Gary is the monoclonal antibody unit manager at the University of Worcester. He also works within a team investigating the development and use of molecular and immunological assay systems to study plant pathogen incidence within horticultural cropping systems.

Many of the projects carried out by this group require the development of specific monoclonal antibodies to selectively recognise a whole host of micro-organisms and antigenic fragments. These include viruses, fungi, bacteria, phytoplasmas, proteins and peptides. Once developed the antibodies can be applied to a number of detection platforms such as enzyme-linked immunosorbent assay, (ELISA), Immunofluorescence, Western blot and lateral flow device technology (LFDs). All of which can be used as an aid in early diagnosis of pathogen presence. Early diagnosis, alongside determination of disease potential re different management practices can result in significant reductions in crop losses.

Teaching

Teaching

Undergraduate teaching:

Gary contributes to teaching modules at the University of Worcester including lectures within the “Infectious agents and allergens” module BIOS 2024. He is also a tutor for the work placement modules BIOS3003, BIOS2003 & EVNS2005.

Areas of expertise:      

Producing primary lepidopteran CpGV (Cydia pomonella granuosis virus) cell lines as well as maintaining, establishing and retaining monoclonal cell lines to a whole host of different antigens.

Working in an Insect Virology group at Warwick HRI (1996-2010), focused on the pathology and molecular biology of lepidopteron and aphid pests by viruses, and the potential of these viruses to be used as bio pesticides.

Carrying out experiments for external companies requiring GLP accreditation. All SOPs had to be generated in-house and continually modified throughout a 3 year period.

Co-founder and Director of Warwick Insect Technologies (WIT) (2005 – 2007), a spinout company from Warwick University specializing in the production and supply of insects and insect relating products which included a technical consultancy service (especially into schools), and the development of alternative entomological applications such as novel silks.

Potential Independent study opportunities:

This group has access to a huge library of monoclonal antibodies to a whole host of different plant diseases. These could be of great benefit to any student who may be interested in learning about different immunological assay systems such as ELISA (enzyme linked immunosorbent assays), lateral flow device technology, and immunofluorescence microscopy. I would be delighted to discuss this further with any potential students who may be interested in developing a project possibly incorporating some of these techniques, towards their Independent studies. These techniques are used to measure antibodies, antigens, proteins and glycoproteins in biological samples.

Examples of previous Independent studies include:

2015       Comparison of DAS ELISA with TAS ELISA for improved detection of Peronospora destructor.

2014       Optimisation of a lateral flow device (LFD) capable of detecting Albugo candida spores.


Research

Research

Current Research Involvement:

  • 2015 – 2018: Agriculture and Horticulture Development Board (£263,999).  Development and testing of single and multiplex diagnostic devices for rapid and precise early detection of Oomycete root and collar rot pathogens for disease avoidance, management and control     
  • 2015 – 2018: Agriculture and Horticulture Development Board (£196,000).  Development and testing of a lateral flow device for both gummy stem blight and powdery mildew in bio-Aerosols during cucurbit production    
  • 2016-2017: Institute of Biomedical Sciences (£4,450).  The development for the rapid diagnosis of the potentially blinding corneal infection Acanthamoeba keratitis

Recent Research Funding Awards:

  • 2016: East Malling Research (£5,219).  “To produce a panel of monoclonal antibodies to “Neonectria ditissima”      
  • 2015: Royal Holloway, University of London (£5,612).  “Maintenance, storage, purification and preliminary kinetic studies of 3 monoclonal antibodies”  

Recent Research Involvement:

  • 2014: Agriculture and Horticulture Development Board (£55,416).  An integrated disease management system to ascribe risk of downy mildew disease on commercial crops in the UK      
  • 2014: Agriculture and Horticulture Development Boar: Extension CP099 (£10,000).
  • 2013: Agriculture and Horticulture Development Board (£67,000).  Clubroot lateral flow in UK commercial Brassica cropping systems
  • 2012: Agriculture and Horticulture Development Board (£122,500).  Lateral flow detection devices for the light leaf spot and powdery mildew vegetable Brassica pathogens and testing of white blister detection test prototypes                         

Publications

Selected Publications

Book chapters:

Wakeham, Alison and Keane, Gary and Proctor, Maureen and Kennedy, Roy (2012) Monitoring Infection Risk for Air and Soil Borne Fungal Plant Pathogens using Antibody and DNA Techniques and Mathematical Models describing Environmental Parameters. In: Microbes in Applied Research: Current Advances and Challenges. World Scientific Publishing Co. Pte. Ltd, Singapore, pp. 152-156. ISBN Paperback: 981-4405-03-5 Ebook: 981-4405-05-8

Journal publications:

Wakeham, A.J.; Keane, G. & Kennedy, R. (2016) Field evaluation of a competitive Lateral-Flow Assay Detection of Alternaria brassicae in vegetable brassica Crops. Plant Disease, 100(9) 1831-1839

Hilton, S.; Bennett, A. J.; Keane, Bending, G.D.; Chandler, D.; Stobart, R. and Mills, P.  (2013) Impact of shortened crop rotation of oilseed rape on soil and rhizosphere microbial diversity in relation to yield decline. PLOS ONE.  8(4):e59859. DOI: 10.1371/journal.pone.0059859

Ryabov, E.V; Keane, G.J; Naish, N.; Evered, C. & Winstanley, D. (2009) Densovirus induces winged morphs in asexual clones of the Rosy apple aphid, Dysaphis plantaginea. Proceedings of the National Academy of Sciences of USA (PNAS), 106 (21): 8465-8470.

Hilton, S.; Kemp, E.; Keane, G. & Winstanley, D. (2008) A Bacmid approach to the genetic manipulation of granuloviruses. Journal of virological methods: 152 (1-2), 56-62

Kingsnorth, C.S; Asher, M.J.C; Keane, G.J; Chwarszczynska, D.M; Luterbacher M.C; & Mutasa-Gottgens, E.S. (2003). Development of a recombinant antibody ELISA test for the detection of Polymyxa betae and its use in resistance screening.  Plant Pathology: 52 (6): 673-680.

Long, S.J.; Richardson, P.N.; Willmott, D.M. and Keane, G.J. (2000) Use of a monoclonal antibody in a field evaluation of the persistence and infectivity of Steinernema n. sp. D1 (Nematoda: Steinernematidae) . Nematology: vol. 2 (4), 425 – 434.

Wakeham, A.J.; Kennedy, R.; Byrne, K. G.; Keane, G & Dewey, F.M. (2000).  Immunomonitoring air-borne fungal plant pathogens: Mycosphaerella brassicicola. EPPO Bulletin (30), 3-4, pp 475-480.

International Conference papers/Abstracts:

Wakeham, A.; Keane, G.; Proctor, M. and Kennedy, R (2011).  Monitoring infection risk for air and soil borne fungal pathogens using antibody and DNA techniques and mathematical models describing environmental parameters.  IV International conference on Environmental, Industrial and applied Microbiology. Biomicroworld, Torremolinos, Malaga, Spain, 14-16th September 2011. Abstracts book p62.

Keane, G.; Asser-Kaiser, S.; Berling, M.; Lopez, M.  Jehle, J. and Winstanley, D. (2008) Comparison of immune responses in Cydia pomonella granulovirus resistant and susceptible strains of C. pomonella. 41st Annual Meeting of the Society for Invertebrate Pathology & 9th International Conference on Bacillus thuringiensis, Univ of Warwick, UK August 3-7th, Abstracts p105   

Ryabov, E; Keane, G; Naish, N. & Winstanley, D. (2008) Pathological effects and possible ecological impact of newly identified viruses of the aphids Brevicoryne brassicae and Dysaphis plantaginea. 41st Annual Meeting of the Society for Invertebrate Pathology & 9th International Conference on Bacillus thuringiensis, Univ of Warwick, UK August 3-7th, Abstracts p121.                                                                

Asser-Kaiser, S.; Winstanley, D.; Keane, G. and Jehle, J. (2008) Investigations on the mechanism of CpGV resistance in Cydia pomonella. 41st Annual Meeting of the Society for Invertebrate Pathology & 9th International Conference on Bacillus thuringiensis, Univ of Warwick, UK August 3-7th, Abstracts p 105.               

Hilton, S.; Keane, G. and Winstanley, D. (2008) Real Time PCR analysis of a mixed infection of granulovirus and nucleopolyhedrovirus from Adoxyphyes orana. 41st Annual Meeting of the Society for Invertebrate Pathology & 9th International Conference on Bacillus thuringiensis, Univ of Warwick, UK August 3-7th, Abstracts p83.

D. Winstanley, G. Cross, N. Naish, G. Keane & S. Hilton (2007). Semichemical autodisemination of tortricid viruses in the field. 40th International Society for Invertebrate Pathology and 1st International Forum for Entomopathogenic Nematodes and symbiotic Bacteria, Quebec, Canada, Aug 12-16th. Abstracts p45. 

D. Winstanley, J.V. Cross, N. Naish, G. Keane, S. Hilton, D. Gajek, R. Van Wese (2005). Laboratory and orchard studies on the transmission of Cydia pomonella granulovirus by contaminated C. pomonella adults. 38th Annual Meeting of the Society for Invertebrate Pathology August 7-11 Anchorage, Alaska, USA

J.V. Cross, D. Winstanley, N. Naish, S. Hilton, G. Keane, R. Van Wezel, D. Gajek. Semichemical driven autodissemination of Cydia pomonella and Adoxyphyes orana baculoviruses. University of Lincoln, Canterbury, New Zealand. 18th January 2005

Cross, J.V., Winstanley, D., Naish, N. F., Hilton, S. L., Keane, G.J.,  Van Wezel, R. &  Gajek, D. (2005) Semichemical driven autodissemination of Cydia pomonella and Adoxyphyes orana baculoviruses. IOBC Bulletin.

J.V. Cross, D. Winstanley, N. Naish, S. Hilton, G. Keane, R. Van Wezel, D. Gajek. Semichemical driven autodissemination of Cydia pomonella and Adoxyphyes orana baculoviruses.. IOBC IFP Baselga di Pine, Italy, 25th – 30th September 2004

D. Winstanley, J.V. Cross, D. Gajek, N. Naish, G. Keane, R. Van Wezel, S. Hilton, Autodissemination of viruses for the control of orchard pests. EMRA subject day, November 4th, 2004, EMR

D. Winstanley, S. L. Wormleaton & G. J. Keane (2002). Recent advances using recombinants of Cydia pomonella granulovirus. XIIth International Congress of Virology. Paris, 27th July - 1st August.

S. L. Wormleaton, G. J. Keane, N. F. Naish & D. Winstanley (2002). A recombinant Cydia pomonella granulovirus expressing green fluorescent protein. XXXV Annual Meeting of the Society of Invertebrate Pathology. Brazil, 18th-23rd August.

Keane, G. & Winstanley, D.; Potential biocontrol agent for the improved control of Cryptophlebia leucotreta. (2001), SIP 34th Annual Meeting, Netherlands, Aug 25th – 30th, Abstract book, p5

Winstanley, D.; Wormleaton, S.L.; Finch, R. & Keane, G.J.  Progress in the genetic improvement of granuloviruses for the control of two important apple pests.  XXI International Congress of Entomology, Brazil 2000, August 20th-26th.   Abstract book 1, p512.

Wakeham, A.J.; Kennedy, R.; Byrne, K.G.; Keane, G. J. & Dewey, F.M. Immunomonitering airborne fungal plant pathogens: Mycosphaerella brassicicola (ringspot).  In: OEPP/EPPO conference on “Diagnostic Techniques for Plant Pests”, Wageningen, 1-4 February, Bulletin 30, 475-480, 2000.

Richardson, P.N.; Keane, G.J.; Willmott, D.M; Long, S.J. & James, B.I. (2000) Monitoring the fate of entomopathogenic nematodes by detecting their symbiotic bacteria. British Crop Protection Council symposium:  Proceedings of an International conference at Brighton, UK, 13-16 November “International Conference on Pests and Diseases “,   (3), 1019 – 1024.

Research contract reports:

WAKEHAM, A., KEANE, G., JOHN, S., & PETCH, G. (2016). Evaluation of an integrated disease management system to ascribe risk of downy mildew disease on commercial salad and bulb onion crops in the UK. HDC CP099C Final Report (47 pages).

WAKEHAM, A., PETTITT, R., KEANE, G., JOHN, S., LEWIS, M. and Edwards, E. (2016) Development and testing of single and multiplex diagnostic devices for rapid and precise early detection of oomycete root and collar rot pathogens for disease avoidance, management and control (40 pages)

KEANE, G. J. & JOHN, S. (2016) To produce a panel of monoclonal antibodies to Neonectria ditissima External contract. (14 pages).

WAKEHAM, A.J., KEANE, G., JOHN, S., PETCH, G. & LEWIS, M. (2015). Development and testing of a lateral flow device for both gummy stem blight and powdery mildew in bio-aerosols during cucurbit production. HDC CP137 Annual Report (29 pages).

WAKEHAM, A.J., LEWIS, M., KEANE, G., EDWARDS, E., PETCH, G.M. & JOHN, S. (2015). Validation of the clubroot lateral flow in UK commercial Brassica cropping systems. HDC CP099 (61 pages)

WAKEHAM, A.J, KEANE, G., JOHN, S., & PETCH, G. (2015). Diagnostics: Validation of the lateral flow detection devices for the light leaf spot and powdery mildew vegetable brassica pathogens and testing of white blister prototypes. HDC CP099a (51 pages).

KEANE, G. J.  (2015) Maintenance, storage, purification and preliminary kinetic studies of 3 monoclonal cell lines (provided). External contract. (12 pages).

WAKEHAM, A.J., LEWIS, M., KEANE, G., PROCTOR, M, PETCH, G. & KENNEDY, R. (2012). Brassicas: Further development of ‘in field’ tests for resting spores of clubroot and the development of clubroot control based on detection. HDC (43 pages).

WAKEHAM, A.J., KEANE, G., PROCTOR, M., PETCH, G. & JOHN, S. & KENNEDY, R. (2012). Onions: Further development and calibration of detection tests for conidia of onion downy mildew in combination with MORPH forecast model milioncast. HDC FV 336 Final report. (29 pages).

WAKEHAM, A.J. & KEANE, G. (2011). Benchmarking predictive models, nutrients and irrigation for management of downy and powdery mildew and white blister. Horticulture Australia Ltd. HAL Final report VG07070. (28 pages)

Presentations/National:

Wakeham, A.J.; Keane, G.J.; John, S.; Townsend, J. & McPherson, M. “Development and testing of diagnostic assays to monitor gummy stem blight and powdery mildew inoculum in air during cucurbit production”. Cucumber and pepper growers’ day at Waltham abbey, UK, 12th October 2016.

Keane, G. Soft Fruit Information day, Presentation on “Deployment of diagnostics in disease management systems for horticulture with emphasis on oomycete stem and root pathogens” The James Hutton Institute, Dundee, Scotland, 19th February 2015.

Trade show/exhibitions/workshops:

UK Onion and carrot conference. “Demonstration on use of onion downy mildew LFD devices” at Kingsgate conference centre, Peterborough, Lincolnshire, 4th November 2015

Winter cauliflower and spring cabbage trials Open day and Conference. “Demonstration of Clubroot lateral flow devices” at Clubroot roadshow at Trevarnon farm, Gwithian, and at Tregenna castle, St. Ives. January 21st 2015 

UK Brassica and Leafy Salad conference and trade exhibition.  “Demonstration of Clubroot lateral flow devices” at Clubroot roadshow, Peterborough, Lincolnshire, Wednesday 28th January 2015

HDC/BGA Brassica technical seminar. “Clubroot update and LFD test demonstration”, Edinburgh 28th January 2014

HDC/Duchy College Brassica Variety Trial conference. “Demonstration of Clubroot lateral flow devices” at Clubroot roadshow, St Ives, 16th January 2014

HDC/BGA Brassica Technical Seminar/workshop. “Practical demonstration of the Clubroot lateral flow device”. Lincolnshire, 14 November 2013