Notes

Slide Show

Outline

1
2	What is UK Biobank?
3	Key principles
4	Protocol development Sample Handling and Storage development sub-group Made recommendations on Samples collected Collection protocols and volumes Cross sectional assays Processing protocols Archiving of sample aliquots Optimal structure of UK Biobank laboratory organization (central vs local processing) Testing and validation of recommended protocol
5	Which sample types should be collected?
6	Which sample types are excluded?
7	Consideration of pre-analytical variables Anticoagulants Haemolysis Clot accelerators Preservatives RNAse Protease inhibitors Fluoride oxalate Borate Azide Cryoprotectants (Central vs local processing) Transport and archiving temperature
8	The following tubes will be used to collect blood from participants
9	A variety of samples will be collected and fractionated
10	The operation will be high throughput, reproducible and standardized…..
11	Samples will be processed centrally:
12	The sample handling pilot Demonstrate that the protocol produces samples that are fit for purpose No general degradation Samples are fit for a wide range of current assays Impact of short term freezing and controlled thawing Impact of delay between sample collection and cryopreservation Is a separate Fluoride Oxalate tube required for measures of plasma glucose concentration?
13	The sample handling pilot - principal
14	Analysis of DNA extracted from white cells and paper media
15	Biochemistry/haematology
16	Biochemistry/haematology
17	EBV immortalization of B-lymphocytes Integration of EBV into B-cell genome via CD21 Existing approaches may require relatively large volumes of blood Cytotoxic T-cell mediated B-cell death may still be a problem (despite use of mitogens and super antigens) Use B-cell selective sorting approach on small volumes of peripheral blood (0.5ml)
18	Distinct stages of B cell maturation
19	EBV immortalization of B-lymphocytes (24 hours)
20	Major findings of sample handling pilot Samples maintained at 4^oC for up to 36 hours prior to processing and cryopreservation are stable although NMR analysis indicates some systematic instability in 36 hour serum Samples maintained at 4^oC for up to 36 hours are suitable for a wide range of analyses Short term freezing and controlled thawing does not impact sample integrity A separate fluoride oxalate tube is not required or measurement of plasma glucose Comparative studies of RNA levels (5’ ends or whole message) will not be possible from these samples
21	Acknowledgements