Details of experiments between Q9BQQ3 and P03126
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ttds-SSRTRREpTQL

Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL

Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 15
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 8
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ttds-SSRTRREpTQL

Average holdup BI: -0.01
Normalized holdup BI: -0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL

Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.06
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.52


Experiment series 5

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL

Average holdup BI: 0.02
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ado-ado-SSRTRRETQL

Average holdup BI: 0
Normalized holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.