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- •Contents
- •Foreword
- •Preface
- •About the Editors
- •Contributors
- •References
- •2.3.4 Barriers to Automation Adoption
- •2.4 Core Ingredients for Successful Digital Transformation
- •2.1 Introduction
- •2.3.1 Operational Challenges
- •2.3.2 Cultural Challenges
- •2.4.2 Cloud Computing
- •2.5 Case Studies of Successful Digital Transformation
- •2.6 Conclusion
- •References
- •3. Computational Protein Design Strategies for Optimization of Antigen Generation to Drive Antibody Discovery
- •3.1 Introduction
- •3.3 Antigen Generation Strategies
- •3.4 Computational Methods
- •3.4.2 Computational Protein Structure Prediction
- •References
- •4. Bioinformatic Analyses of Antibody Repertoires and Their Roles in Modern Antibody Drug Discovery
- •4.1 Introduction
- •4.6 Summary and Future Directions
- •Acknowledgments
- •References
- •5.1 Introduction
- •5.2 Databases
- •5.2.1 Databases in Machine Learning Approaches
- •5.2.2 Database Types
- •5.3 Applications of Machine Learning in Antibody Discovery and Development
- •5.3.1 Structure Prediction with Deep Learning
- •5.3.3 Developability
- •5.4 Antibody Generation and Design by Language Models
- •5.4.1 Antibody Representations
- •5.4.2 Representation Learning
- •5.4.3 Language Models
- •References
- •6.1 Introduction
- •6.2 Antibody Generation through Deep Generative Models
- •6.3.1 Sampling and Scoring
- •6.5 Conclusions and Perspectives
- •Acknowledgments
- •References
- •7.1 Introduction
- •7.2.3 Computational Approaches to Predict Antibody–Antigen Interaction
- •7.3 Conclusion
- •Competing Interests
- •Acknowledgments
- •References
- •8.2 Common Types of Molecular Simulations for Biomolecules
- •8.2.1 Molecular Dynamics (MD) Simulations
- •8.2.2 Monte Carlo (MC) Simulations
- •8.2.3 Challenges of Molecular Simulations
- •8.3.1 Periodic Boundary Conditions
- •8.4 Uses of Molecular Simulation in Antibody Drug Development
- •8.5 Conclusion
- •References
- •9. Considerations of Developability During the Early Stages of Antibody Drug Discovery and Design
- •9.1 Introduction
- •9.2 Historical Perspective
- •9.3 Clinical Antibody Data Set
- •9.5 Control Antibodies
- •9.7 Assessment of Chemical Liabilities
- •9.8 Conclusions and Future Perspectives
- •Acknowledgments
- •References
- •Abbreviations
- •10.1 Introduction
- •10.4.1 Conclusions and Outlook
- •Acknowledgments
- •References
- •11.8 Conclusions and Future Directions
- •References
- •12.1 Introduction to PK/PD and QSP Modeling
- •12.1.1 PK/PD Modeling
- •12.1.2 QSP Modeling
- •12.2.1 Monoclonal Antibodies (mAbs)
- •12.2.3 Cell Therapies
- •12.2.4 Gene Therapies
- •12.2.5 Vaccines
- •12.2.6 mRNA/siRNA/Oligonucleotide Therapeutics
- •12.4 Case Studies
- •12.5 Conclusions and Future Perspectives
- •References
- •13.1 Introduction
- •13.2 AI/ML: A Game Changer for Antibody Design
- •13.3 Multispecific Antibody Design
- •13.4 Adapting AI to the Design of Multispecific Antibodies
- •13.4.1 Structure Prediction and Modeling
- •13.4.2 Developability Prediction and Optimization
- •13.4.4 In Silico Modeling and Simulation
- •13.5 The Future: Beyond Optimization
- •13.5.1 Market Trends and Commercialization
- •13.5.2 Logic Gates, Biosensors, and De Novo Design
- •13.5.3 Challenges and Opportunities
- •13.6 Conclusion
- •Acknowledgments
- •References
- •Index

5 • Articial Intelligence and Machine Learning 101
5.4.3.3 Transformer architectures
Recently, transformer architectures have been gaining popularity in the representation
learning area as they achieved state‑of‑the‑art results on a wide range of NLP tasks
(Wolf etal. 2020). Using self‑attention mechanisms, transformers are able to learn rela‑
tionships between words in sentences and, therefore, produce distributed representa‑
tions. Such neural networks are often trained with the MLM approach in which one
masks or changes part of the input and the model learns to predict the altered part.
5.4.3.4 ProtBERT
To this time, there have been several adaptations of transformer architectures to antibody
data. For example, Elnaggar and colleagues (Elnaggar etal., n.d.) showed that embeddings
obtained from transformers captured relevant biological information as small‑size mod‑
els trained solely on those representations were able to compete with bigger architectures
on various tasks such as protein classication. Their neural networks reached compa‑
rable performance to methods utilizing MSA, which indicates that transformer‑produced
embeddings could be a good starting point for various downstream tasks.
5.4.3.5 AbLang
This transformer architecture was also successfully applied in the eld of antibody
representation by Olsen and colleagues (Olsen etal. 2022b) who trained the AbLang
model on antibody sequences from OAS. The resulting architecture consists of two
parts: AbRep and AbHead, which produce representations for sequences and predict the
probability of each amino acid on all positions, respectively. Pre‑training was based on
the RoBERTa approach (Liu etal. 2019). The team showed the biological information
encoded into the vectors by drawing 10,000 naïve and 10,000memory B‑cell sequence
representations (Ghraichy etal. 2021) using t‑SNE and compared the results to evo‑
lutionary scale modeling 1b (ESM‑1b) embeddings (Rives etal. 2021). Both models
could separate antibody sequences by their V gene families, but AbLang yielded better
separation of naïve and memory B cells. The resulting transformer model was capable
of restoring missing residues in immunoglobulin sequences, obtaining similar or better
results than using IMGT germlines, but without the knowledge of the germlines.
5.4.3.6 AntiBERTa
As proposed by Leem and colleagues (Leem etal. 2022), AntiBERTa (Antibody‑specic
Bidirectional Encoder Representation from Transformers) is another example of trans‑
former architecture. The model was pre‑trained using the RoBERTa approach on 57mil‑
lion human BCR sequences from 61 studies available in OAS. The team selected random
1000BCR heavy‑chain sequences (Ghraichy etal. 2021) from naïve and memory B‑cell
sequences and showed that on the top of mutational load and V gene used, embeddings
carry information about B‑cell type–they were able to partition naïve and memory B‑cell
sequences in the representation space. This result was compared to embeddings prepared

102 Biopharmaceutical Informatics
by ProtBERT (Elnaggar etal. 2020), where the separation was not as clear, which indi‑
cates that compared to the general protein transformer model, AntiBERTa representa‑
tions encapsulate more antibody‑specic information. Leem and colleagues noticed that
high self‑attention scores presented residue pairs of contacts indicating that the model
is capable of understanding the structural information. So, they applied it to the para‑
tope prediction problem, where the model classied each residue from the input sequence.
This was achieved by adding a classication head on top of the already existing 12lay‑
ers. The prediction results were compared to Parapred (Liberis etal. 2018) and ProABC
(Olimpieri etal. 2013), which demonstrated SoTa results in paratope prediction. The team
used the model to produce embeddings of known therapeutic antibodies, showing that it
was possible to determine their origin (human, murine, humanized, chimeric) as well as,
to a certain degree, the correlation with immunogenicity response scores–ADA. This
demonstrates that embeddings learned biologically relevant information, and the learned
representations correspond to B‑cell origin, immunogenicity, and structure.
5.4.3.7 AntiBERTy
AntiBERTy, another model based on the BERT architecture, was proposed by Ruffolo
and colleagues (Ruffolo etal. 2021). It was trained on 558million sequences from OAS
with MLM objective. The team analyzed repertoires from donors with HIV‑1 neutral‑
izing VRC01 antibodies. For each sample, they created a k bearers neighbour (kNN)
graph using model embeddings and visualized it in two‑dimensions using Uniform
Manifold Approximation and Projection (UMAP). Using these plots, they observed tra‑
jectories from germline sequences and mutated derivatives corresponding to sequence
changes in the afnity maturation process. With repertoire data, individual sequences
are not labeled. Hence–relying on clonal expansion–the team produced noisy labels,
and frequently observed sequences were assumed to be binders. Next, they applied mul‑
tiple instance learning (MIL) to predict whether the sets of sequences contain binding
antibodies. They created single‑instance bags of sequences from known VRC01 anti‑
bodies, conrming that the model produces the correct positive predictions. Finally,
they annotated each antibody structure with attention, and in most cases (7 out of 10
sequences), attention‑pointed binding residues.
5.4.3.8 AbBERT
Another transformer architecture called AbBERT trained on 20million heavy and light
sequences from OAS was published by Vashchenko and colleagues (Vashchenko etal.
2022). The model family is based on ProtBERT, but ne‑tuned on antibodies. Both
heavy and light sequences were used to train the models, during which the team anno‑
tated functional regions of input sequences by inserting additional annotation tokens,
before and after all CDRs. First, the authors showed that the model is capable of predict‑
ing CDR regions. The team introduces the term “humanness” that is used to measure
the similarity between input immunoglobulin sequence and antibodies sampled from
people. This score was used to evaluate 600 known therapeutic sequences that have
passed various clinical trials and showed that antibodies with a low assigned metric tend

5 • Articial Intelligence and Machine Learning 103
to be immunogenic. Then, the model was applied for in silico antibody optimization in
which the baseline anti‑SARS‑CoV‑1 antibody sequence was modied so that the opti‑
mized immunoglobulin was able to bind to another target–SARS‑CoV‑2. Model with
AbBERT embeddings on input was used to solve the optimization problem. Finally, they
performed in vitro experiments which demonstrated that poorly scored sequences were
weakly expressed in the cells. They have also observed a correlation between the model
scores and the protein stability metrics calculated using Free Energy Perturbation.
5.4.3.9 BioPhi (Sapiens module)
Sapiens is one of the two BioPhi models aiming at antibody humanization. Similarly,
to other SoTa models, it is a transformer‑based model trained toward the MLM goal.
Two separate models for light and heavy chains have been created, each having 568,857
parameters and being based on the RoBERTa model. The training dataset consisted of
human‑only and unaligned sequences from OAS: 20million heavy and 19million light
sequences. Analysis of the averaged attention matrix showed high importance between
CDR loops, which are close structurally but apart in sequence–thus proving that the
model is able to recognize long‑range interactions.
Antibody humanization works by leveraging the fact that only human mAbs were
used for training. Input variable region sequence is processed by the model, giving prob‑
abilities for all 20 amino acids for all positions. The most probable residues for frame‑
works are selected, keeping unchanged CDRs from input, which minimizes the risk of
affecting binding properties but making antibodies more similar to human ones. Such a
procedure has been performed on 177 antibodies (25 with known parental sequence and
152 humanized mAbs with presumed original sequence), obtaining results comparable
to human experts.
5.5 CONCLUSIONS AND FUTURE
PERSPECTIVES IN AI FOR
ANTIBODY DISCOVERY
Over the past 40 years, antibodies have rmly established their role as the most important
group of biologics. Up until now, the development of currently 100 approved antibody
therapeutics relied on a “discovery” process driven by experimental laboratory‑based
methods. Thanks to advances in high‑throughput experimental data generation as well
as progress in computational model development, it is possible to shift the paradigm
from antibody discovery toward “design.”
For designing a novel biologic computationally, one requires two elements. First,
one needs to generate biologically or physically plausible sequences and structures.
Second, one requires objective functions to gauge whether the molecule has the proper‑
ties expected of it. The sampling of novel molecules has been greatly facilitated by gen‑
erative modeling such as variational auto encoders (VAEs), GANs, and language models

104 Biopharmaceutical Informatics
that learn the representation of antibodies from large‑scale NGS data. The performance
of predicting objective antibody features such as antibody‑antigen binding or develop‑
ability still needs to be addressed. Nevertheless, researchers have started to combine the
two features, generating molecules that are either biased or ltered for those with better
biophysical features.
As such, computational methods are now capable of producing naturally viable
starting points that are free from statistically obvious liabilities. Achieving the goal of
fully computational antibody design–as opposed to “discovery”–still requires improv‑
ing the prediction of the molecule’s therapeutic features, chiey binding and develop‑
ability. On the binding front, one could hope for a modeling revolution, on par with
structure prediction as the two problems bear many parallels. However, on the develop‑
ability front, hoping for such progress is fanciful, mostly due to the lack of data.
Developability is an umbrella term uniting multiple biological assays. Even though
many of these are regularly performed at organizations developing biologics, such a
plethora of data was scarcely envisaged for training models. Therefore, data are often‑
times not comparable between different runs, projects, and teams since they were
generated with a specic therapeutic challenge rather than to develop a generalistic
developability prediction method. For this reason, a new paradigm emerges called “pre‑
diction‑rst,” where data are generated specically with model training in mind. Over
the short term, they might not contribute to any therapeutic projects, but rather act as
a long‑term investment into the development of a foundation for a broadly applicable
computational model.
All in all, shifting from discovery to design and from project‑driven data genera‑
tion toward prediction‑rst requires a sizable shift within the organizations responsible
for biologics development. Understandably, it is a large diversion of resources from
well‑proved experimental methods to the development of innovative methods that still
need to be validated. Nevertheless, with the ongoing progress in the development of
computational models for antibodies, such a shift has become far more realistic.
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