Honestly the only reason I noticed was I went to tweak my seat height! Never used anything other than a torque wrench either 😔

Shieeet. Well hopefully tweaking contact points can get you out there shredding

Biggest tires you can, tubeless at lowish pressure. Made a huge difference for me (herniated disc + sciatica)

Huge thanks to co-authors Hayley Knox @basicchemist.bsky.social & Brian Cho, mentors Barbara Imperiali & Karen Allen, and everyone who made this possible!

Still amazed this worked so well. Check out the full preprint and let me know what you think!

#StructuralBiology #YeastDisplay #Science

Why should you care? This enables:
🔬 CryoEM of challenging membrane proteins
💊 Better membrane protein therapeutics
🧪 Fluorescent probes for cell biology
📊 Structure-based drug design
All without detergent artifacts!

We also demonstrated our nanobodies could be functionalized via sortase mediated ligation, allowing for the design of bespoke molecular probes!

We validated binding through multiple methods:

- Biolayer interferometry
- Size exclusion chromatography
- Native SMA-PAGE

The Nb-protein complexes remained stable even through SEC - perfect for structural studies! 💪

What makes this special?
✅ Proteins stay in native-like lipid environment
✅ Works with picomole amounts of protein
✅ No fluorescent labeling artifacts
✅ Produces stable, high-affinity binders

But here's the real beauty - the binding curves! Our selected nanobodies show high affinity binding with KD values from 15-200 nM. 🎯

The magic happens during selection! Look at this beautiful enrichment of WbaP binders over successive rounds:

From 0.7% → 18% positive binders! Even protein concentrations as low as 100 nM

First, we proved the purification method is generalizable. We successfully purified 5 different bacterial membrane proteins in SMALPs:

- Different sizes (26-94 kDa)
- Various topologies
- Different oligomeric states

All worked! 💪

Here's our selection strategy! We use the same dual-Strep tag for both purification AND magnetic bead capture. No protein modification needed!

The workflow: Negative sorts → Positive selection → Amplify → Repeat 🔄

The challenge: Traditional nanobody selection requires either llama immunization 🦙 or detergent-solubilized proteins. But detergents can distort membrane protein structure!
Our solution? Keep proteins in polymer-stabilized liponanoparticles (SMALPs) throughout selection

Huh, I thought they were out of iceland?