BIOMARKER tracker matrix continuation — lysosomal markers + aSyn / GlcSph / SCSF assay rows, Annapurna note, C8H CSF LCMS, Dr Margreith UMCS oligo aSyn discussion, MID/ELISA Parkin assay (cGAMP / pSer65-Ub), CSF Buffer 125 pg/mL row + Migdalisa Richards Ciegg patient sample notes

(This page is a dense biomarker development tracker grid that runs across many narrow color-coded cells. The full per-row content is preserved in format_notes; key visible rows are reproduced below.)

BIOMARKER tracker — lysosomal markers + aSyn

rowcontent (visible)
ARM lysosomal markersHox 70, Lamp1 & 2nx Lamp2 in disease model (PD) and animal model (By WB) (BM discovery) sample 가급 Eslau Sugita for ph.
Direction of lysosomal markerchanges may be affected by disease stage of PD; Beclin이 증가해서 proteolysis가 정상적이면, 치료로 이거 낮추면 무슨 의미?
lysosomal (marker) changes in a broader context
global protein degradation

GlcSph (Sato)

20210904 Otake Roche needs to be used MAFA. (1F0 ECEN Pooled MJFF in plasma low Otake Annapurna no found that this assay value is the flash report in the coming week. Initially I want to test if the LCSF is detectable or not within May. PV. We need further qualifications and may make conclusion. CSF was found in highly prioritized to PMM PI #15 just alternative MN, no real prioritization is scheduled in May. If you want to acclimate it, let’s talk with Otake-san…

What does it detect?: capture most of the existing α-synuclein forms in CSF? (total α-synuclein), to quantify total amounts of the protein and not disease-specific isoforms or the post-translationally modified and aggregated forms of the protein.

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most of the antibodies displayed in commercial immunoassay for total αSYN target the C-terminal domain of αSYN(Figure 1), which is known to harbor the majority of αSYN PTMs, including phosphorylation nitration and ubiquitination [38,39]. It is possible that these PTMs could either eliminate or mask the epitope of these antibodies or interfere with antibody binding to αSYN
measured in CSFnormal levels

GlcSph & GalSph (Sato) — assay status

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20210414 OtakeNo difference HC vs PD [‘As in PD MCl’ vs HC PCl] PV15 aSyn is measurable in human CSF and plasma; The concentration is approximately 10x higher in plasma than in CSF
Data from PD samples to be sent to BAH on Jan 24 vs HCs status it does not have significantly clinical meaning between elevated levels of α-syn in CSF on PD continued and may potentially be a synaptic neurodegeneration in 1 context
SatoDGGM (in human serum plasma & CSF)
20210706Pooled HC CSF & all human plasma 0.05 ng/ml in plasma all serum 0.07 ng/ml as a way recovery next 0.05 ng/mL Po PV ng/mL; for the assay (Otake-san: Annapurna 0.4 mg/ml is reasonable based on Praxis’s report)
20211211 Otakethe conditions for the assay value initialization start cyno serum + Pooled in cyno plasma 0.05 ng/ml in plasma all serum 0.07 ng/ml ic-cell aSyn was developed and PD samples can be analyzed

C8H CSF LCMS

Timeline by Disease clusterby 178 / 2020 / 1Q
If we can develop quantification at least it highly reproducible qualitative assay through collaboration with Dr Hatano BIA can be achieved by Q4 FY20
20210714 OtakeThis is the assay value to optimize a quantitative method PD’s data is available based on Q1 FY21

CSF (LCMS) — Annapurna SOMA / Juntendo collaboration

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20210715In the collaboration with Juntendo Univ, we want to evaluate to what extent the assay can be improved as quantitative samples for PA reproducibility / FY20 to 4Q FY21
20211211 Otakesubstrate (in soluble monomers a/syn) to be sourced from large amounts in the optimization ‘cross-site validation’ tech transfer reproducibility (K negative) validation in Parkin PD Model
20210803 SteveThere is no current plan to resurrect it. The ADx group has some very high affinity antibodies from MJFF to develop an assay and every worked on recombinant oligomeric standards. But when testing on human CSF, the signal was below the LOQ and they hypothesis was that the amount of oligomeric in CSF was too low. That’s why the RT-QuiC or PMCA requires a long incubation to accumulate the oligomers and rely on the self-aggregation of the protein to amplify the signal.

Dr Margreith UMCS Steve (USPS) wide cluster row

No (Disease cluster)by 178 / 2020 / 1Q
GBA Project T1 70
But aSyn HD Of Stickly’s leading vending samples 22 SCT Tr15 14 PD aSyn (in PD/Hyposmia) 30 Roche
Roche Eli oligo aSyn (Boys CSF aSyn Roche aSyn 22’M /11/Lamb of (Roche))

cGAMP / Parkin / Substantia / pSer65-Ub assay rows

transcript factor91 / 134 / 250 / 020 SEP / High sensitive cGAMP measurement assay based on LCMS / Verification of the cGAMP quantification using CSF and mice brain and blood by LCMS and immunoassay / 2020 / 2021 Q3 / Verification using patient sample
MID or ELISA candidates Akram etc2020 / 02 / 1Q / 0 / mBD (LcGCAS-G2-CB) Cd5 / accuracy and precision / detectability of pasit in human CSF / concentration of psasit in human CSF (1) / leagues to be ready 202012
20240105MJ assay method development by MQ ASM Ay Feasibility assessment will be performed by end Q3
20210624MJ method dev for parking protein in ongoing Material on successfully selected highly sensitive (around 30pq/ml LOQ) Ab pair for parking and will conduct assay validation enrichment method is currently ongoing Preliminary results showed detectable signal with a sample enrichment process This result will be carefully verified
20210906Preliminary work of customized MMIA assay indicated 0.6 pg/ml in buffer based (LcQ probably is 50pl that is 40 times higher than that in customized MMIA assay reported last month Human CSF samples will be measured early next month
cGAMP Sample will be analyzed using spike recovery and dilution linearity are confirmed

CSF Buffer 125 pg/mL row

Buffer125 (pg/mL)24.4 (pg/mL)0.61
Scatter plot legendPARK2sPDHC

Migdalisa Richards Ciegg

Verification using patient sample. Generated data can be valid in any comparison of relative Parkin Level among diagnoses. However interpolated value (the pg/ml) should be handled as tentative.

20120114 Currently we only have one PD CSF sample but we can evaluate parkin levels using both the SoEL and 50ML of human CSF.

20210210 Capture parkin antibody (anti-Parkin antibody, kfsutra) ABcam, rabbit (#PR3PA7-376). Detection antibody anti-Parkin antibody (Abcam, rabbit (#PR3PA7-214)).