🚨 My lab is hiring at all levels!

Interested in animal origins & evolutionary cell biology?

I'm recruiting a postdoc, PhD students & a research assistant to study the molecular evolution of cell adhesion using marine invertebrates + comparative genomics.

🔗: clarkelab.com/join/

Please repost!

Peak New England.

#Boston #massacusetts #MA

Adventure day!

#gravelbike #bike #MA

QOTSA Boston night two was WILD

#QOTSA #boston

I live with a criminal

#dog

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

Well that's not great 💀

Honestly jealous of Ashley's new Seigla 🤩

#gravelbike #lauf

This guy was collecting tolls on the morning ride

#gravelbike #biking #massachusetts
#boston

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)

Vibes:

🐶
#dog

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! 💪

A workflow for selecting binders against membrane proteins in liponanoparticles

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

Selection of nanobodies against liponanoparticle-embedded membrane proteins by yeast surface display Single-domain antibodies, known as nanobodies (Nbs), are widely used in structural biology, therapeutics, and as molecular probes in biology and biotechnology. Nbs towards soluble proteins are routine...

🧵
Excited to share our latest preprint!

www.biorxiv.org/content/10.1...

We developed a method to generate high-affinity binders against membrane proteins in their native-like environment!

🧪 #MembraneProteins #Nanobodies

Huh, I thought they were out of iceland?

Lauf?

Guilty

Got it for an absolute steal when GT announced the "production pause". Its such a fun ride!

Cinturato Ms looking sick on the gravel rig. The jump from 40 to 45mm makes a huge difference in ride quality!

#Gravel #GravelBike #cycling #bike #pirelli #GT

Whelp looks like I'm sleeping on the couch now