Carbohydrate-deficient transferrin determined in blood microsamples from healthy newborns by using capillary zone electrophoresis.
OBJECTIVE
There is a paucity of studies on quantitative determination of carbohydrate-deficient transferrin (CDT) in newborns. The aim of our study was therefore to determine CDT concentrations in newborns by using capillary zone electrophoresis (CZE).


METHODS
Capillary blood was collected from the heels of 28 at-term healthy newborns, simultaneously with the Guthrie card screening. Forty-seven adults were examined as controls. CZE separations were performed with a P/ACE MDQ capillary electropherograph in uncoated fused-silica capillaries using a commercial reagent kit. After iron saturation, the samples were loaded by application of 0.5 psi for 15 s, and separated under 28kV with UV detection. All relevant transferrin (Tf) glycoforms were separated within 7 min. CDT quantification (%CDT) was carried out by calculating the percentage ratio between the sum of the peak areas of CDT-related glycoforms and the sum of peak areas of all Tf glycoforms.


RESULTS
In most cases, good separations of Tf glycoforms were obtained. In the newborns the %CDT was 0.51 versus 0.66 in adults (difference not statistically significant). Trisialo-Tf concentration was significantly lower in newborns (3.20) than in adults (4.11). Furthermore, pentasialo-Tf appeared to be lower in newborns (7.30) than in adults (14.00), but because complete separation of the peaks of tetrasialo- and pentasialo-Tf was not always possible, this finding could not be confirmed statistically.


CONCLUSIONS
CZE showed definite advantages in terms of volume of blood to be collected, simplicity and standardization of analysis and, because of the direct detection of the separated zones, accuracy of quantification. The present study provides the basic information in the search for glycosylation defects in newborns.
Zamboni
G
G
Department of Mother and Child, Biology-Genetics, Section of Paediatrics, University of Verona, Verona, Italy. Giorgio.Zamboni@univr.it
Bortolotti
F
F
Zaffanello
M
M
De Paoli
G
G
Tagliaro
F
F
D000328:Adult; D015415:Biomarkers / Q000097:blood; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008832:Microchemistry / Q000379:methods*; D008875:Middle Aged; D013695:Temperance; D014168:Transferrin / Q000031:analogs & derivatives* / Q000032:analysis
D015415:Biomarkers; D014168:Transferrin; C056759:carbohydrate-deficient transferrin