Supplemental TableI.Reagents and buffers used in the various Gyrolab assays

Assay / Modality / Capture (Biotin) / Detector (Alexa Fluor 647) / Assay Buffer / Wash 1 Buffer / Wash 2 Buffer / Species/ Matrix
1 / ADC / Gt anti-hu IgG; Southern Biotech (cat# 2049-08) / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / A / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Mk plasma
2, 3 / ADC / Ms anti-payload;
internal reagent / Dk anti-hu IgG (H+L; Jackson Immunoresearch
(cat# 709-605-149) / B / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Rt and Mk plasma
4 / MAb / Dk anti-hu IgG (H+L); Jackson Immunoresearch (cat# 709-065-149) / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / D / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Rt plasma
5, 6 / MAb or Monovalent MAb / Gt anti-hu IgG; Bethyl Laboratories
(cat# A80-419B) / Gt anti-hu IgG (H+L); Molecular Probes
(cat# A-21445) / E / PBS, 0.01% Tween-20 / 20% ETOH, 0.5% SDS / Ms diluted blood
7 / MAb / Gt anti-hu IgG; Southern Biotech
(cat# 2049-08) / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / C / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Ms diluted blood
8 / MAb / Dk anti-hu IgG (H+L); Jackson Immunoresearch (cat# 709-065-149) / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / D / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Rt plasma
9 / ADC / Ms anti-payload;
internal reagent / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / F / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Ms diluted blood
10 / ADC / Gt anti-hu IgG; Southern Biotech
(cat# 2049-08) / Dk anti-hu IgG (H+L); Jackson Immunoresearch
(cat# 709-605-149) / F / PBS, 0.01% Tween-20 / 50mM Glycine, 0.5% SDS, 20% ETOH, pH 9.5 / Ms diluted blood
11 / ADC / Gt anti-hu IgG; Southern Biotech (cat# 2049-08) / Gt anti-hu IgG (H+L) Bethyl Laboratories (cat# A80-319A) labeled internally / G / PBS, 0.05% Tween-20 / 20% ETOH, 0.5% SDS / Ms diluted blood
12 / ADC / Gt anti-hu IgG; Southern Biotech (cat# 2049-08) / Ms or Rb anti-payload antibody;
internal reagent / G / PBS, 0.05% Tween-20 / 20% ETOH, 0.5% SDS / Ms diluted blood
13-15 / ADC / Gt anti-hu IgG; Southern Biotech (cat# 2049-08) / Ms anti-hu kappa (SB81a); Southern Biotech (cat#9230-01)
labeled internally / H / PBS, 0.1% Tween-20 / 20% ethanol, 1.5M NaCl / Mk and Rt plasma or serum
16-19 / ADC / Sh anti-hu IgG (H+L); The Binding Site
(cat# AU003.MCUS01) / Gt anti-hu IgG (H+L) Bethyl Laboratories (cat# A80-319A) labeled internally / G / PBS, 0.1% Tween-20 / 20% ETOH, 0.5% SDS / Mk and Rt plasma or serum
20-23 / ADC / Sh anti-hu IgG (H+L); The Binding Site
(cat# AU003.MCUS01) / Ms or Rb anti-payload antibody;
internal reagent / G / PBS, 0.1% Tween-20 / 20% ETOH, 0.5% SDS / Mk and Rt plasma or serum

A 1:1 v/v SuperBlockT20 (PBS) : 0.2M Tricine, pH 8.5, 0.1% Tween-20, 0.15M NaCl, 0.05% Proclin 300.

B Buffer A with added 100 ug/ml mouse IgG.

C 0.2M Tricine, pH 8.5, 1% BSA, 0.15M NaCl, 0.1% Tween-20, 0.05% Proclin300.

D 0.2M Tricine, pH 8.5, 1% BSA, 0.5M NaCl, 0.1% Zwittergent, 0.05% Proclin300.

E Rexxip A Buffer (Gyros, Inc., Upsala, Sweden, cat# P0004820).

F 0.1M HEPES, 0.15M NaCl, 3 mM EDTA, 0.005% surfactant P20, 1% w/v BSA.

G Super Block (Scytek Laboratories, West Logan, UT, cat# AAA125).

H StartingBlock T20 (Thermo Fisher Scientific, Pittsburgh, PA, cat# EN37539).

PBS = phosphate buffered saline. ETOH = ethanol. SDS = sodium dodecyl sulfate.

Dk = donkey, Gt = goat, Hu = human, Mk = monkey, Ms = mouse, Rb = rabbit, Rt = rat and Sh = sheep.

1