Mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders caused by
Mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders caused by the deficiency of lysosomal enzymes. with the same molecular weights can be separated by liquid chromatography. We have also developed GPR120 modulator 2 another GAG assay by high-throughput mass spectrometry (HT-MS/MS). The HT-MS/MS consists of a solid phase extraction robot that binds and de-salts samples from assay plates and directly injects them into a MS/MS detector, reducing sample processing time to within ten mere seconds. HT-MS/MS as a result yields much faster throughput than standard LC-MS/MS-based methods; however, the HT-MS/MS system does not make use of a chromatographic step, and therefore, cannot independent GAGs that have the same molecular weights. Both techniques can be applied to the analysis of dried blood spots, blood, and urine specimens. With this review, we describe the assay methods for GAGs and the application to newborn testing and analysis of MPS. 1(6S)4GlcNAc(6S) (31, 32), and, consequently, this ELISA approach does not quantify total KS. Furthermore, the ELISA GPR120 modulator 2 method cannot detect less than 2.5 ng/mL of KS, whereas LC-MS/MS GPR120 modulator 2 can measure as little as 0.2 ng/mL of KS. As a result, in rodents that synthesize far less KS, blood levels are measurable by LC-MS/MS but not by standard ELISA. Dermatan Sulfate (DS) All MPS VI individuals (n = 4) showed an elevation of plasma DS. MPS I (18/22, 81.8%), MPS II (26/27, 96.3%), and MPS VII (2/7, Mouse monoclonal to CD4.CD4 is a co-receptor involved in immune response (co-receptor activity in binding to MHC class II molecules) and HIV infection (CD4 is primary receptor for HIV-1 surface glycoprotein gp120). CD4 regulates T-cell activation, T/B-cell adhesion, T-cell diferentiation, T-cell selection and signal transduction 28.6%) individuals had a significant elevation of DS as well. These findings show that DS measurements by LC-MS/MS are applicable to the testing for most MPS I, II, and VI individuals (18). Heparan Sulfate (HS) All MPS I, II, and III individuals (n = 60) showed a significant elevation of plasma DiHS-0S and DiHS-NS. The group of MPS III individuals comprised five IIIA, four IIIB, and two IIIC individuals. Two out of 6 individuals experienced a significant elevation of HS (18). The results showed 1) that blood levels of DiHS-NS and DiHS-0S were significantly elevated in individuals with MPS II and III, 2) that individuals with a severe form of MPS II experienced a higher level of HS than those with an attenuated form, and 3) that reduction of blood HS was seen in MPS II individuals treated with ERT or HSCT (18, 19). Composition of DS and HS in Blood The compositional percentage of DiHS-0S, DiHS-NS, and Di-4S GPR120 modulator 2 in blood samples of MPS individuals was compared. The ratio of each GAG composition was expected to be affected by deficiency of a specific enzyme. In the normal control samples, the percentage of DiHS-0S, DiHS-NS, and Di-4S was 40.4%, 7.7%, and 51.9%, respectively. The proportion of Di-4S was higher in individuals with MPS VI, compared to that in normal settings (mean; 80.6% 51.9%). The proportion of DiHS-0S was higher in individuals with MPS III and VII, compared to that in normal settings (mean; 56.4% 40.4%; 65.1% 40.4%). The proportion of DiHS-NS was also higher in MPS III individuals compared to that in normal settings (mean; 19.7% 7.7%). Other types of MPS did not provide any difference in ratios of DS and HS (18, 25). 3.4. Newborn MPS It is critical to elucidate when GAGs start to accumulate in cells of individuals to determine feasibility of measuring GAGs for newborn screening for MPS. To evaluate whether the LC-MS/MS method can distinguish MPS newborns from healthy control newborns, we assayed DS and HS levels in DBS samples that had been obtained at birth from six individuals later diagnosed with MPS (four MPS I, one MPS II, and one MPS VII). All six instances showed elevations of DS and HS levels, compared with those of control newborns (26). HT-MS/MS also shown that the levels of DiHS-0S and DiHS-NS are elevated in DBS acquired at birth from 11 individuals diagnosed with MPS I (n = 6) or MPS III (n = 5), when compared to control newborn DBS (19). The levels of DiHS-0S and DiHS-NS from DBS of a newborn with MPS II were 3 and 1.5 times higher than in DBS from control newborns. With this study DiHS-0S was more discriminating than DiHS-NS in separating individuals from settings. Ruijter have also demonstrated that.