SBIR-STTR Award

Development of a Lipoprotein Analyzer system is proposed for rapid analysis of and characterization of the HDL, LDL, and VLDL profiles of human blood plasma. Preliminary results using a prototype system demonstrated the potential of the proposed system: the HDL, and VLDL fractions were clearly separated, plasma samples were analyzed without sample pre-treatment, and differences in lipoprotein profiles were noted for different patients. Analyses times were between 20 minutes and 1 hour. Determination of accurate particle size of the lipoprotein complexes also demonstrated the unique capabilities of this system. The separation strategy of the proposed Lipoprotein Analyzer is based on Flow field-flow fractionation, (Flow FFF) an elution based technique. This methodology is capable of high resolution measurements and is bio-compatible for analysis of biological macromolecules ranging from proteins up to chromosomes and cells. Thus, this instrumentation is potentially a powerful and effective tool for rapid and direct measurement of the entire subfraction set of the lipoprotein profile. The applicants propose to further study, optimize, and develop Flow FFF to ultimately generate a rugged, routine technique. They expect this development will provide a more expedient and less expensive method for lipoprotein characterization than currently available.Awardee's statement of the potential commercial applications of the research: The need for screening serum cholesterol levels is immense, and so there is a large market potential for the Lipoprotein Analyzer. The technique is expected to provide a rapid, low cost alternative to the current techniques. Also, the system could be used in research laboratories for direct determination of lipoprotein particle size and for semi-preparative fractionation of the lipoprotein component.

Thesaurus Terms:
biomedical equipment development, blood fractionation, blood lipoprotein, clinical biomedical equipment, physical separation high density lipoprotein, low density lipoprotein, very low density lipoprotein National Heart, Lung and Blood Institute (NHLBI)

Further development of a Lipoprotein Analyzer based on flow field-flow fractionation (Flow FFF) is proposed. Flow FFF is an innovative technology capable of high resolution separations. This methodology is gentle, flexible and biocompatible for analysis of biological macromolecules ranging from proteins to cells. The driving force for separation is generated via application of an external field -- in this case, a cross flow field. External application allows the separation to be easily tuned for desired resolution / analysis time. Thus, Flow Fff is a powerful and effective tool for rapid, direct measurement of each subfraction of the lipoprotein profile. Phase I work successfully demonstrated separation of HDL, LDL, and VLDL components in human blood plasma. Recovery rates were comparable or better than that of traditional ultracentrifugal technique. Analysis times were on the order of 20 minutes per sample. System cost < $30,000. Phase II is directed at further demonstrating accuracy and precision, increasing sample throughput, and streamlining system design / operation for user convenience. The overall result of this work will be an inexpensive direct technique for simultaneous quantitation of HDL, LDL, and VLDL components in human blood plasma. Patient fasting will not be required as triglyceride levels are not involved in the measurement. PROPOSED COMMERCIAL APPLICATION: Current techniques for screening serum cholesterol are indirect or otherwise inadequate. The proposed system will provide a rapid, direct, low cost alternative to current techniques. Patient fasting will not be required. The specifications & design of the proposed system considers commercial needs of lipoprotein research & centralized clinical labs as well as needs for increased precision, accuracy, & reliability.

Thesaurus Terms:
biomedical automation, biomedical equipment development, blood fractionation, blood lipoprotein