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Reshaping Antibody Selection: Early Characterization of Large mAb Panels

Revamps Lead Identification and Technology Development

Reshaping Antibody Selection: Early Characterization of Large mAb Panels

Revamps Lead Identification and Technology Development

Zimple Matharu and Christine BeeBristol-Myers Squibb Redwood City

Carterra User Group Meeting2019-09-10

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Antibody generation at BMS

Mouse strains Routes of immunization

People

Immunogens

Antibodytechnologies

Dosingregimen

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LSA assays for selection

Capture kinetics

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LSA assays for selection

Coupled Ab kinetics and binning

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Comparison of HT kinetics

kd(1

/s) c

oupl

edAb

s, L

SA

kd (1/s) captured Abs. LSAka

(s) c

oupl

edAb

s, L

SAka (s) captured Abs. LSA

KD(M

) cou

pled

Abs,

LSA

KD (M) captured Abs, LSA

KD (M) captured Abs, Biacore

KD(M

) cap

ture

dAb

s, L

SA

ka (s) captured Abs, Biacore

ka(s

) cap

ture

dAb

s, L

SA

kd(1

/s)

capt

ured

Abs.

LSA

kd (1/s) captured Abs. Biacore

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Binning

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Affinities and epitope communities

120pM

120nM

nd (>125nM)

Low affinity samples can cause errors.

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Replication in groups of related samples

120pM

120nM

nd (>125nM)

Curated binning shows identical behavior of highly related samples.

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Redundancy within communities

related samples, same selection technology, same bin

High redundancy within sample subset→ change selection criteria

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Mouse strains and epitope communities

Much higher diversity observed in red mouse strain.

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Mouse strains and affinities by epitope

120pM 120nM nd (>125nM)

Different mouse strains correlate with different affinities.Red mouse strain produces higher affinities and higher diversity.

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Capturing full diversity in sequence and epitope

Multiple sequence alignment by bin:• harness full diversity by combining

sequencing and binning• choose low sequence liability

upfront• QC binning results

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Capturing full diversity

Multiple sequence alignment by bin:• harness full diversity by combining

sequencing and binning• choose low sequence liability

upfront• QC binning results

Expanded by epitope mapping, cell binding, functional data, biophysical and biochemical characterization,…

Combined with affinities:• understand affinity maturation• evaluate and understand performance of transgenic mouse strains• explore epitope specific binding characteristics• inform immunization and selection technologies

Pick an optimal panel of diversified leads

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Acknowledgements

Haibin ChenGinger RakestrawKe LiMatt TomlinsonGabriel WuJulian LowLore Florin

Flavio SchwarzAndy DengLuhua ZhangRenu JainRalston BarnesGavin Dollinger

Carterra

Sumetha KannanFaisal MalikJohn MajercakErin O’BrienMike HornsbyArvind Rajpal