MACS Matchmaker

Anti-tag capture adapters — protocol

The lino anti-tag adapter system uses oriented capture proteins (Protein A/G + Protein A/G/L, trisNTA, Strep-Tactin XT, anti-GFP VHH) that are pre-conjugated to a chosen oligonucleotide sequence and ship ready-to-use from the catalogue. Each adapter hybridises to the Oligo|PEG chip via DDI, then the tagged ligand is loaded in a capture step. The chip is regenerable at the duplex level between different tagged ligands, and the captured ligand can additionally be eluted via its tag (e.g. imidazole for His-tag, low pH for Fc-captured IgGs) without stripping the duplex.

Available adapters

All catalogue adapters share the same format: 110 µL stock at 2.6 µM in PBST, enough for 10 immobilisations at the recommended 200 nM working concentration (130 µL/injection). Each is pre-conjugated to a single Seq ID (00–16) chosen at order time — combine adapters with different Seq IDs to multiplex on a standard 8×8 Oligo|PEG chip (6×9 on request).

AdapterTag / ligand capturedElutionWebshop
Protein A/G + Protein A/G/LHuman / mouse IgG (Fc region) for the A/G variant; A/G/L also captures kappa light chains (Fabs, scFvs, Vκ nanobodies)pH 2.0 glycine or duplex stripCHF 195
TrisNTA6×His-tag (requires NiSO₄ pre-loading — not included)250 mM imidazole or 50 mM EDTACHF 195
Strep-Tactin XTStrep-tag II and Twin-Strep-tag5 mM desthiobiotin or duplex stripCHF 195
Anti-GFP VHHGFP-fusion proteinsDuplex stripCHF 195

General protocol (all adapters)

Each adapter follows the same three-step DDI capture workflow:

  1. Adapter-oligo hybridisation. Flow the adapter-oligo conjugate (200 nM in PBST) over the Oligo|PEG chip for 5 min. The chip is now loaded with oriented capture sites.
  2. Tagged ligand capture. Flow the tagged protein at a sub-saturating concentration (typically 10–100 nM) for 3–5 min. Monitor the signal in real time and stop when the desired loading level is reached.
  3. Analyte measurement. Run the analyte concentration series at 30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch) for association and 200 µL/min for dissociation. Regenerate with tag-specific elution or strip the duplex for a fresh adapter load.

Protein A/G — IgG and antibody fragment capture

ParameterValue
AdapterProtein A/G-oligo (Fc capture) or Protein A/G/L-oligo (Fc + Vκ capture) — order one Seq ID per loaded ridge
Adapter stock110 µL at 2.6 µM in PBST (10 immobilisations per vial)
Working dilution200 nM in PBST, 130 µL injection
Hybridisation flow rate / contact time10 µL/min, 5 min
IgG loading concentration10–100 nM in PBST
Loading flow rate / contact time10 µL/min, 3–5 min (to desired signal, not necessarily saturation)
SCK association flow rate30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch)
SCK dissociation flow rate200 µL/min
Regeneration option A (tag elution)10 mM glycine pH 2.0, 30 s — releases IgG, leaves adapter on chip
Regeneration option B (duplex strip)3 M guanidinium-HCl + 125 mM NaOH, 400 µL/min, 0.5 min

TrisNTA — His-tag capture

ParameterValue
AdapterTrisNTA-oligo, one Seq ID per loaded ridge
Adapter stock110 µL at 2.6 µM in PBST (10 immobilisations per vial)
Working dilution200 nM in PBST, 130 µL injection
Hybridisation flow rate / contact time10 µL/min, 5 min
Ni²⁺ pre-loading500 µM NiSO₄ in PBST (NiSO₄ not included in the adapter vial), 10 µL/min, 2 min; wash with PBST (200 µL/min) to remove excess Ni²⁺ before loading His-tagged protein
His-tagged ligand concentration10–100 nM in PBST
Loading flow rate / contact time10 µL/min, 3–5 min
SCK association flow rate30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch)
SCK dissociation flow rate200 µL/min
Regeneration option A (His elution)250 mM imidazole in PBST — releases His-tagged protein
Regeneration option B (chelation)50 mM EDTA in PBST — strips Ni²⁺ and ligand; re-load Ni²⁺ to reuse
Regeneration option C (duplex strip)3 M guanidinium-HCl + 125 mM NaOH, 400 µL/min, 0.5 min

Strep-Tactin XT — Strep-tag II capture

ParameterValue
AdapterStrep-Tactin XT-oligo, one Seq ID per loaded ridge
Adapter stock110 µL at 2.6 µM in PBST (10 immobilisations per vial)
Working dilution200 nM in PBST, 130 µL injection
Hybridisation flow rate / contact time10 µL/min, 5 min
Strep-tagged ligand concentration10–50 nM in PBST
Loading flow rate / contact time10 µL/min, 3–5 min
SCK association flow rate30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch)
SCK dissociation flow rate200 µL/min
Regeneration (tag elution)5 mM desthiobiotin in PBST, 2 min — releases Strep-tag II ligand; Strep-Tactin XT adapter remains on chip for next capture
Regeneration (duplex strip)3 M guanidinium-HCl + 125 mM NaOH, 400 µL/min, 0.5 min

Practical notes

See also: immobilisation method comparison table.