Control cell populations for detection of paroxysmal nocturnal hemoglobinuria
AssigneeBiolegend, Inc.
InventorsSusannah Helene Kassmer-Rodriguez, Anagha Ashok Divekar, David Pierre Michel Pillon
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Abstract
Provided are systems and methods that may be employed to detect the presence or absence of paroxysmal nocturnal hemoglobinuria (PNH) in a subject that includes a leukocyte including GPI-anchors the leukocyte incubated with one or more blocking agents each independently specific to one or more GPI-anchored proteins, the leukocyte further incubated with a pro-aerolysin or derivative thereof, and optionally a second leukocyte including GPI-anchors not incubated with one or more of the blocking agents. The systems may be used as a positive control in assays detecting the presence or absence of PNH.
Claims20
A system configured for use in detection of the presence or absence of paroxysmal nocturnal hemoglobinuria comprising:
- a first leukocyte comprising GPI-anchors, said first leukocyte incubated with one or more blocking agents each independently specific to one or more GPI-anchored proteins;
- and
- said leukocyte further incubated with a pro-aerolysin or derivative thereof.
A process of detecting the presence or absence of paroxysmal nocturnal hemoglobinuria (PNH) in a subject comprising:
- obtaining a first sample of whole blood from said subject, said sample optionally lysed, fixed, and washed;
- contacting said sample with a blocking solution comprising one or more blocking antibodies each independently specific to one or more GPI-anchored proteins and a pro-aerolysin or derivative thereof, and incubating for a first incubation time to form a blocked sample;
- combining said blocked sample with a PNH antibody cocktail;
- and
- subsequently subjecting said blocked sample to analysis by flow cytometry to detect the presence or absence of paroxysmal nocturnal hemoglobinuria in said subject.
The process of 7, wherein the one or more blocking antibodies to one or more GPI-anchored proteins is specific for CD157, CD24, or CD14.
A process of analyzing a positive control for paroxysmal nocturnal hemoglobinuria (PNH) comprising:
- obtaining a leukocyte comprising GPI-anchors, said leukocyte incubated with one or more blocking antibodies each independently specific to one or more GPI-anchored proteins;
- further incubating said leukocyte with a pro-aerolysin or derivative thereof to form a blocked leukocyte;
- further incubating said blocked leukocyte with a PNH antibody cocktail;
- combining the blocked leukocyte with an unblocked leukocyte to form a leukocyte mix;
- and
- subjecting said leukocyte mix to analysis by flow cytometry.
Dependencies are inferred from the claim text.