Chymotrypsin treatment of HLA-A and B molecules reveals 1D-IEF variation

Susceptibility to birdshot chorioretinopathy is restricted to the HLA‐A29.2 subtype

“…The exact mechanism by which the presence of The bands at B8 level are partially digested B44.1 (B8 after enzymatic treatment has the same isoelectric point as B44.1 without treatment). The shift in band position following chymotrypsin treatment is consistent with the loss of the cytoplasmic domain from the tyrosine 321 of the HLA heavy chain (6). Papain treatment gave the same pattern (data not shown).…”

“…I allowed us to determine a resolution of 0.03 pH unit/mm of gel in the HLA-A29.2 area. At this resolution, two HLA molecules different by a single substitution of a charged amino acid would have been distinguished according to their migration pattern (6,11).…”

Comprehensive, serologically equivalent DNA typing for HLA‐B by PCR using sequence‐specific primers (PCR‐SSP)

“…Serology is a quick and convenient method of HLA class I detection but it is hindered in many cases by serological cross-reactivity and a lack of useful typing reagents. One-dimensional isoelectric focusing (1,2) can define class I polymorphisms but is realistically used only as a supplement to serology in order to define additional subtypes. To improve HLA class I typing DNA-based typing methods have been developed such as PCR followed by hybridisation with sequence-specific probes (3.4.5).…”

Identification of HLA‐B35, B53, B18, B5, B78, and B17 alleles by the polymerase chain reaction using sequence‐specific primers (PCR‐SSP)

“…The panel members were typed for HLA class I antigens by serology and included 14 B35, 5 B53, 4 B18, 8 B51, 2 B52, 2 B78, 12 B57 and 2 B58 positive individuals. Some panel donors were also typed by ID-IEF as previously described (2).…”