US 11,747,271 B2
Triple-internal standard based glycan structural assignment method for capillary electrophoresis analysis of carbohydrates
Gabor Jarvas, Veszprem (HU); Marton Geza Szigeti, Budapest (HU); Andras Guttman, Brea, CA (US); and Jeff Chapman, Santa Ana, CA (US)
Assigned to DH Technologies Development Pte.Ltd., Singapore (SG)
Appl. No. 16/328,208
Filed by DH TECHNOLOGIES DEVELOPMENT PTE. LTD., Singapore (SG)
PCT Filed Aug. 17, 2017, PCT No. PCT/IB2017/054989
§ 371(c)(1), (2) Date Feb. 25, 2019,
PCT Pub. No. WO2018/037316, PCT Pub. Date Mar. 1, 2018.
Claims priority of provisional application 62/518,104, filed on Jun. 12, 2017.
Claims priority of provisional application 62/380,062, filed on Aug. 26, 2016.
Prior Publication US 2019/0234867 A1, Aug. 1, 2019
Int. Cl. G01N 21/33 (2006.01); G01N 33/53 (2006.01); G16C 20/20 (2019.01); G01N 21/64 (2006.01); G01N 27/447 (2006.01); G01N 30/04 (2006.01); G01N 30/88 (2006.01)
CPC G01N 21/33 (2013.01) [G01N 33/5308 (2013.01); G16C 20/20 (2019.02); G01N 21/6402 (2013.01); G01N 27/44726 (2013.01); G01N 2030/045 (2013.01); G01N 2030/8836 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A system for identifying the structure of a glycan by co-injecting with the glycan a triple internal standard made up of three different glucose α1-4 oligomers of maltooligosaccharide (MOL), comprising:
a separation device that receives a known or unknown glycan that is co-injected with three different oligomers of MOL and separates the glycan and the three different oligomers of MOL over time, wherein MOL comprises oligomers that have a degree of polymerization (DP) ranging from unity up to dozens, wherein the ratio of the difference in migration time between two consecutive oligomers of MOL and the difference in migration time between adjacent consecutive oligomers of MOL is a first known constant value in a first region of the DP range of MOL and a second known constant value in a second region of the DP range of MOL, and wherein two of the three different oligomers of MOL are selected to produce migration times in one of the first region or the second region and the third of the three different oligomers of MOL is selected to produce a migration time in the other of the first region or the second region;
a detector that measures the separated known or unknown glycan and the separated three different oligomers of MOL as intensity peaks that are a function of migration time; and
a processor that
receives the intensity peaks of the separated known or unknown glycan and the separated three different oligomers of MOL,
calculates the migration times of a plurality of other oligomers of MOL from the migration times of the three different oligomers of MOL producing a virtual ladder of migration times for oligomers of MOL and the first known constant value and the second known constant value,
calculates glucose unit (GU) values for the intensity peaks of the separated known or unknown glycan by comparing their migration times to the calculated migration times of the virtual ladder of migration times for oligomers of MOL, and
identifies the structure of the known or unknown glycan by comparing the calculated GU values of the intensity peaks of the separated known or unknown glycan to a database of GU values for known glycan structures.