Supplementary Materials

Supplementary Material for:

Urine lipoarabinomannan glycan in HIV-negative patients with pulmonary tuberculosis correlates with disease severity

Luisa Paris, Ruben Magni, Fatima Zaidi, Robyn Araujo, Neal Saini, Michael Harpole, Jorge Coronel, Daniela E. Kirwan, Hannah Steinberg, Robert H. Gilman, Emanuel F. Petricoin, Roberto Nisini, Alessandra Luchini,* Lance Liotta

*Corresponding author. Email: aluchini{at}gmu.edu

Published 13 December 2017, Sci. Transl. Med. 9, eaal2807 (2017)
DOI: 10.1126/scitranslmed.aal2807

This PDF file includes:

  • Materials and Methods
  • Fig. S1. The Kd affinity between RB221 and LAM exceeds that of FB28.
  • Fig. S2. Copper dyes outperform copper free dyes such as fast blue B and safranin O.
  • Fig. S3. Nanocages dissociate biomarker from interfering substances, in silico mathematical modeling.
  • Fig. S4. CS-35 mAb is specific for LAM diluted in human urine, batch verification.
  • Fig. S5. Competition assay confirmed the specificity of CS-35 mAb.
  • Fig. S6. Coupling chemistry to covalently incorporate the FB28 dye in the inner volume of the nanocages.
  • Fig. S7. LAM binding to RB221 and depletion from supernatant are independent of pH in a 5 to 7 range.
  • Fig. S8. RB221 binding to LAM is hindered by the presence of a copper-chelating agent (EDTA).
  • Fig. S9. RB221-LAM interaction requires intact diol moieties of LAM as proven by NaIO4 oxidation.
  • Fig. S10. Carbohydrate concentration in the LAM reference standard (0.160 mg/ml) was quantified by a linear colorimetric assay.
  • Fig. S11. Plot of the 95% CI of the sensitivity and specificity of the ROC analysis reported in Fig. 3C.
  • Fig. S12. The RB221 dye is immobilized in the inner volume of the cages and is available for high–molecular weight ligand binding after cross-link degradation and consequent increase of the effective pore size.
  • Fig. S13. CS-35 anti-LAM mAb does not cross-react with purified polysaccharides from N. meningitidis and S. pneumoniae.
  • Fig. S14. Nanocage capturing followed by CS-35 antibody detection is specific for LAM and does not cross-react with M. tuberculosis lipomannan and arabinogalactan.
  • Table S1. Nanocage bait chemistries screened to capture and enrich LAM from human urine.
  • Table S2. Medical characteristics of diseased TB-negative controls.
  • Table S3. Urinalysis results for all study participants.

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